Bioactive polypeptides for improvements in plant protection, growth and productivity

ABSTRACT

Bioactive priming polypeptides are provided that are useful when applied to plants in agricultural formulations. Methods of using the formulations containing the bioactive priming polypeptides are also provided which are applied exogenously to the surface of a plant or a plant cell membrane or endogenously to the interior of a plant or to a plant cell. The bioactive priming polypeptides when applied to a plant, a plant part, or a plant growth medium or a rhizosphere in an area surrounding the plant or the plant part increase growth, yield, health, longevity, productivity, and/or vigor of a plant or a plant part and/or decrease abiotic stress in the plant or the plant part and/or protect the plant or the plant part from disease, insects and/or nematodes, and/or increase the innate immune response of the plant or the plant part and/or change plant architecture.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Provisional Application No.62/534,710, filed Jul. 20, 2017, the content of which is incorporatedherein by reference in its entirety.

FIELD OF THE INVENTION

Bioactive priming polypeptides are provided which can be delivered inagricultural formulations. The polypeptides can be applied to crops toachieve agronomically desirable outcomes such as enhanced phenotypes inplants (e.g., those that exhibit protection against pest, disease agentsand abiotic stress), increased plant growth, productivity and yield.

BACKGROUND OF THE INVENTION

Conventional methods to achieve desired agronomic phenotypes such asincreased yield, disease prevention, disease resistance, and improvedabiotic stress tolerance have utilized mostly selective breeding,grafting, transgenic and agrochemical approaches.

Bioactive Priming Polypeptides Involved in Plant Defense Responses

Plants possess an immune system that detects and protects againstmicrobes that can cause disease. Antimicrobial peptides (AMPs) in plantsare often the first line of defense against invading pathogens and areinvolved in the initiation of defense responses that can impart innateimmunity to a plant. Many AMPs are generically active against variouskinds of infectious agents. They are generally classified asantibacterial, anti-fungal, anti-viral and/or anti-parasitic.

The resistance of given plant species against certain pathogenicorganisms that can contact a plant surface and colonize it, is based onhighly specialized recognition systems for molecules produced only bycertain microbes (for example, specific bacterial or fungal strains).Plants sense potential microbial invaders by using pattern-recognitionreceptors (PRRs) to recognize the pathogen-associated molecular patterns(PAMPs) associated with them.

Flagellin/Flagellin-Associated Polypeptides

Flagellins and flagellin-associated polypeptides derived from thoseflagellins have been reported primarily to have functional roles ininnate immune responses in plants. These polypeptides are derived fromhighly conserved domains of eubacterial flagellin. Flagellin is the mainbuilding block of the bacterial flagellum. The flagellin protein subunitbuilding up the filament of bacterial flagellum can act as a potentelicitor in cells to mount defense-related responses in various plantspecies.

“Flagellin” is a globular protein that arranges itself in a hollowcylinder to form the filament in a bacterial flagellum. Flagellin is theprincipal substituent of bacterial flagellum, and is present inflagellated bacteria. Plants can perceive, combat infection and mountdefense signaling against bacterial microbes through the recognition ofconserved epitopes, such as the stretch of 22 amino acids (Flg22)located in the N-terminus of a full length flagellin coding sequence.The elicitor activity of Flg22 polypeptide is attributed to thisconserved domain within the N-terminus of the flagellin protein (Felixet al., 1999). Plants can perceive bacterial flagellin through a patternrecognition receptor (PRR) at the plant's cell surface known asflagellin sensitive receptor, which is a leucine-rich repeat receptorkinase located in the plasma membrane and available at the plant cellsurface. In plants, the best-characterized PRR is FLAGELLIN SENSING 2(FLS2), which is highly conserved in both monocot and dicot plants.

In Arabidopsis, the innate immune response to Flg22 involves a hostrecognition protein complex that contains the FLS2 leucine rich repeat(LRR) receptor kinase (Gómez-Gómez L. and Boller T., “FLS2: An LRRreceptor-like kinase involved in the perception of the bacterialelicitor flagellin in Arabidopsis,” Molecular Cell 5: 1003-1011, 2000).In Arabidopsis thaliana, FLS2 is a PRR that determines flagellinperception and is specific for the binding of the flagellin-associatedpolypeptide(s). For example, the binding of Flg22 to the outer plantFLS2 membrane-bound receptor triggers a signaling cascade that isinvolved in the innate immune response that induces the plant to mount ahighly specific signaling-associated cascade that is involved in theactivation of pattern-triggered immunity (Chinchilla et al., “TheArabidopsis receptor kinase FLS2 binds Flg22 and determines thespecificity of flagellin perception,” Plant Cell 18: 465-476, 2006).Thus, the binding of Flg22 to the Arabidopsis FLS2 membrane-boundreceptor promotes the first step of activation in which the bindingelicits an activation cascade for defense responses in the plant. TheFlg22-FLS2 interaction can also lead to the production of reactiveoxygen species (ROS) that contribute to the induction of an oxidativeburst, cellular medium alkalinization, downstream induction ofpathogen-responsive genes and defense-related responses which then canimpart disease resistance to a plant (Felix G. et al., “Plants have asensitive perception system for the most conserved domain of bacterialflagellin,” The Plant Journal 18: 265-276, 1999, Gómez-Gómez L. andBoller T., “FLS2: An LRR receptor-like kinase involved in the perceptionof the bacterial elicitor flagellin in Arabidopsis,” Molecular Cell 5:1003-1011, 2000, Meindi et al., “The bacterial elicitor flagellinactivates its receptor in tomato cells according to the address-messageconcept,” The Plant Cell 12: 1783-1794, 2000). In tomato, high affinitybinding of Flg22 to a FLS receptor was observed using both intact cellsas well as to microsomal membrane preparations. In this study, thebinding of Flg22 to the FLS2 receptor(s) at the plasma membrane surfacewas nonreversible under physiological conditions, which reflects anuptake process of the Flg22 elicitor with import into the tomato cells(Meindi et al., “The bacterial elicitor flagellin activates its receptorin tomato cells according to the address-message concept,” The PlantCell 12: 1783-1794, 2000). Recognition of Flg22 by FLS2 triggers bothlocal and systemic plant immune responses. The Flg22-bound, activatedFLS2 receptor complex is internalized into plant cells by endocytosisand moves systemically throughout the plant (Jelenska et al., “Flagellinpeptide flg22 gains access to long-distance trafficking in Arabidopsisvia its receptor, FLS2,” Journal of Experimental Botany 68: 1769-1783,2017), which may contribute towards systemic Flg22 immune responses.

Flagellin receptor perception mediation involving Flg22 is highlyconserved across divergent plant taxa (Taki et al., “Analysis offlagellin perception mediated by flg22 receptor OsFLS2 in rice,”Molecular Plant Microbe Interactions 21: 1635-1642, 2008). Submicromolarconcentrations of synthetic polypeptides comprising between 15-22 or 28amino acids from conserved domains of a flagellin protein, act aselicitors to initiate defense responses in a variety of plant species.

Generation of transgenic plants has been used to confirm theflagellin-specific PAMPs that bind to the flagellin-specific PRRs.Ectopic expression of FLS2 in Arabidopsis plants showed a directcorrelation between the flagellin responses and FLS2 expression levels,which indicate that FLS2 is involved in the recognition of flagellin (asignal of bacterial presence) and leads to the activation of defenseresponses in plants (Gómez-Gómez L. and Boller T., “FLS2: An LRRreceptor-like kinase involved in the perception of the bacterialelicitor flagellin in Arabidopsis,” Molecular Cell 5: 1003-1011, 2000).Transgenic plants expressing the flagellin binding receptor have shownefficacy against certain pathogens. Flagellin binding to FLS2 wasinvolved in the initiation of expression of specific MAP kinasetranscription factors that function downstream of the flagellin receptorFLS2. Mutant plants (fls2) lacking in the FLS2 receptor are insensitiveto Flg22 (Gómez-Gómez L. and Boller T., “FLS2: An LRR receptor-likekinase involved in the perception of the bacterial elicitor flagellin inArabidopsis,” Molecular Cell 5: 1003-1011, 2000), and impaired in Flg22binding to the FLS2 receptor. Mutant plants (fls2) also exhibitedenhanced susceptibility to infection and disease when treated withpathogenic bacteria (Zipfel et al., “Bacterial disease resistance inArabidopsis through flagellin perception,” Nature 428: 764-767, 2004).

Traditionally, methods to improve disease resistance have capitalized onthese and other such findings and have taken a transgenic approach.Transgenic plants and seeds transformed with a Flagellin-Sensing (FLS)receptor protein (WO2016007606A2 incorporated herein by reference in itsentirety) or with transcription factors involved in downstream signalingof FLS (WO2002072782A2 incorporated herein by reference in its entirety)have produced plants that confer disease resistance to certainpathogenic microorganisms. In another example, transgenic plantsexpressing Flagellin-Sensing (FLS3) receptor also have exhibitedenhanced resistance to disease compared to non-transgenic plants notexpressing the FLS3 receptor (WO2016007606A2 incorporated herein byreference in its entirety).

Plant Defensins/Thionins

Plant defensins are also characterized as anti-microbial peptides(AMPs). Plant defensins contain several conserved cysteinyl residuesthat form disulphide bridges and contribute to their structuralstability. Defensins are among the best characterized cysteine-rich AMPsin plants. Members of the defensin family have four disulfide bridgesthat fold into a globular structure. This highly conserved structurebestows highly specialized roles in protecting plants against microbialpathogenic organisms (Nawrot et al., “Plant antimicrobial peptides,”Folia Microbiology 59: 181-196, 2014).

Thionins are cystine-rich plant AMPs classified in the defensin familyand typically comprise 45-48 amino acid residues, in which 6-8 of theseamino acids are cysteine that form 3-4 disulfide bonds in higher plants.Thionins have been found to be present in both monocot and dicot plantsand their expression can be induced by infection with various microbes(Tam et. al., “Antimicrobial peptides from plants,” Pharmaceuticals 8:711-757, 2015). Particular amino acids of thionins such as Lys1 andTyr13, which are highly conserved, have been found to be vital to thefunctional toxicity of these AMPs.

Harpin and Harpin-Like (HpaG-Like)

Similar to the flagellins or the flagellin-associated polypeptides,harpins comprise a group of bacterial-derived elicitors that are derivedfrom larger precursor proteins. Harpins are critical for the elicitationof a hypersensitive response (HR) when infiltrated into theintercellular space or apoplast of plant cells (Kim et al., “Mutationalanalysis of Xanthomonas harpin HpaG identifies a key functional regionthat elicits the hypersensitive response in nonhost plants,” Journal ofBacteriology 186: 6239-6247, 2004). Application of the distantharpin-like (HpaG-like) bioactive priming polypeptide(s) to a plantprovides an alternative conduit to protect a plant from disease andinsect pressure. Harpins utilize a type III secretion system that enablethe transport of proteins across the lipid bilayers that makeup theplant plasma cell membrane. The binding of harpins to the surface of theplasma cell membrane can trigger an innate immune response thatresembles those triggered by pathogen-associated molecular patterns(PAMPs) and are known to activate PAMP-triggered immunity (Engelhardt etal., “Separable roles of the Pseudomonas syringae pv. phaseolicolaaccessory protein HrpZ1 in ion-conducting pore formation and activationof plant immunity,” The Plant Journal 57: 706-717, 2009). Mutationalanalysis of a harpin-like HpaG derived polypeptide showed that the 12amino acid residues between Leu-39 and Leu50 of the original 133 aminoacid harpin elicitor precursor protein was critical to the elicitationof a hypersensitive (HR) and subsequent innate immune responses intobacco (Kim et al., “Mutational analysis of Xanthomonas harpin HpaGidentifies a key functional region that elicits the hypersensitiveresponse in nonhost plants,” Journal of Bacteriology 186: 6239-6247,2004). This indicates that a specific amino acid region of harpins(similar to the other AMPs) is responsible for the elicitationresponses. Harpins, such as HpaG-like can be used to enhance resistanceto not only plant pathogens but also to insects (Choi et al., “Harpins,multifunctional proteins secreted by gram-negative plant pathogenicbacteria,” Molecular Plant Microbe Interactions 26: 1115-1122, 2013).Harpin has been used to induce disease resistance in plants and protectplants from colonization and feeding by insect phloem-feeding insects,such as aphids (Zhang et al., “Harpin-induced expression and transgenicoverexpression of phloem protein gene At.PP2A1 in Arabidopsis repressphloem feeding of the green peach aphid Myzus persicae,” BMC PlantBiology 11: 1-11, 2011).

Elongation Factor Tu (EF-Tu)

Elongation factor Tu is an abundant protein found in bacteria and actsas a pathogen-associated molecular pattern (PAMP) to initiate signalingcascades that are involved in plant disease resistance and plant innateimmunity to microbial pathogenic organisms. Interestingly, some EF-Tupolypeptides are also found to exist in plants. The first 18 amino acidresidues of the N-terminus of EF-Tu from Escherichia coli, termed elf18,is known to be a potent inducer of PAMP-triggered immune responses inplants (Zipfel et al., “Perception of the bacterial PAMP EF-Tu by theReceptor EFR restricts Agrobacterium-mediated transformation,” Cell 125:749-760, 2006). Polypeptides derived from E. coli EF-Tu are perceived bythe plant cell-surface localized receptor EF-Tu receptor (EFR) (Zipfelet al., 2006). EF-Tu binding and activation of EFR follow a similar modeof action compared to that of the Flg peptide-FLS2 receptor complex(Mbengue et al., “Clathrin-dependent endocytosis is required forimmunity mediated by pattern recognition receptor kinases,” Proc NatlAcad Sci U.S.A. 113: 11034-9, 2016).

Growth Altering Bioactive Priming Polypeptides

Phytosulfokines (PSKα)

Phytosulfokines (PSK) belong to a group of sulfated plant polypeptidesthat are encoded by precursor genes that are ubiquitously present andhighly conserved in higher plants (Sauter M., “Phytosulfokine peptidesignaling,” Journal of Experimental Biology 66: 1-9, 2015). PSK genesare encoded by small gene families that are present in both monocots anddicots and encode a PSK polypeptide(s) that can be active as either apentapeptide or a C-terminally truncated tetrapeptide (Lorbiecke R,Sauter M, “Comparative analysis of PSK peptide growth factor precursorhomologs,” Plant Science 163: 348-357, 2002).

The phytosulfokine protein is targeted to the secretory pathway inplants by a conserved signal polypeptide (Lorbiecke R, Sauter M,“Comparative analysis of PSK peptide growth factor precursor homologs,”Plant Science 163: 348-357, 2002). Processing of the phytosulfokineprecursor protein involves sulfonylation by a tyrosylproteinsulfotransferase within the plant secretory pathway, specifically thetrans-Golgi followed by secretion and proteolytic cleavage in theapoplast in order to produce PSK (Sauter M., “Phytosulfokine peptidesignaling,” Journal of Experimental Biology 66: 1-9, 2015). After PSK isprocessed from the larger precursor polypeptide, the polypeptideundergoes tyrosine sulphation (Ryan et al., “Polypeptide hormones,” ThePlant Cell Supplement, S251-S264, 2002). The secreted polypeptide isthen perceived at the cell surface by a membrane-bound receptor kinaseof the leucine-rich repeat family (Sauter M., “Phytosulfokine peptidesignaling,” Journal of Experimental Biology 66: 1-9, 2015 where PSK canthen bind to the specialized PSK receptor (for example, PSK1 fromArabidopsis) which has a leucine-rich repeat region located on the plantplasma membrane surface. Specific binding of PSK was detected in plasmamembrane fractions from cell suspension cultures derived from rice andmaize and the binding to the receptor was shown to initiate andstimulate cell proliferation (Matsubayashi et al., “Phytosulfokine-α, asulfated pentapeptide, stimulates the proliferation of rice cells bymeans of specific high- and low-affinity binding sites,” ProceedingsNational Academy of Science USA 94:13357-13362, 1997).

Phytosulfokines (PSK) serve as sulfated growth factors with biostimulantactivities and are involved in the control of the development of rootand shoot apical meristems, growth regulation and reproductiveprocesses. PSKs have also been reported to initiate cell proliferation,differentiation of quiescent tissues and are involved in the formationand stimulation and differentiation of tracheary elements (Matsubayashiet al., “The endogenous sulfated pentapeptide phytosulfokine-αstimulates tracheary element differentiation of isolated mesophyll cellsof zinnia, Plant Physiology 120: 1043-1048, 1999). PSK signaling hasalso been reported to be involved in the regulation of root andhypocotyl elongation that occurs in Arabidopsis seedlings (Kutschmar etal., “PSK-α promotes root growth in Arabidopsis,” New Phytologist 181:820-831, 2009).

Root Hair Promoting Polypeptide (RHPP)

Root hair promoting polypeptide (RHPP) is a 12 amino acid fragmentderived from soybean Kunitz trypsin inhibitor (KTI) protein, which wasdetected from soybean meal that was subjected to degradation using analkaline protease from Bacillus circulans HA₁₂ (Matsumiya Y. and Kubo M.“Soybean and Nutrition, Chapter 11: Soybean Peptide: Novel plant growthpromoting peptide from soybean,” Agricultural and Biological Sciences,Sheny H. E. (editor), pgs. 215-230, 2011). When applied to soybeanroots, RHPP was shown to accumulate in the roots and promote root growththrough the stimulation of cell division and root hair differentiationin Brassica.

SUMMARY OF THE INVENTION

A polypeptide is provided for bioactive priming of a plant or a plantpart to increase growth, yield, health, longevity, productivity, and/orvigor of a plant or a plant part and/or decrease abiotic stress in theplant or the plant part and/or protect the plant or the plant part fromdisease, insects and/or nematodes, and/or increase the innate immuneresponse of the plant or the plant part and/or change plantarchitecture. The polypeptide comprises either:

(a) a flagellin or flagellin-associated polypeptide and an amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 1-225, 227-375, 526, 528, 530, 532, 534,536, 538, 540, 541, 751 and 752; or

(b) a mutant flagellin or flagellin-associated polypeptide and an aminoacid sequence of the mutant flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 571-579 and 753; or

(c) a mutant flagellin or flagellin-associated polypeptide and an aminoacid sequence of the mutant flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 580-586; or

(d) a retro inverso Flg22 polypeptide and an amino acid sequence of theretro inverso Flg22 polypeptide comprises any one of SEQ ID NOs:376-450, 527, 531, 533, 535, 537 and 539; or

(e) a retro inverso FlgII-28 polypeptide and an amino acid sequence ofthe retro inverso FlgII-28 polypeptide comprises any one of SEQ ID NOs:451-525; or

(f) a retro inverso Flg15 polypeptide and an amino acid sequence of theretro inverso Flg15 polypeptide comprises SEQ ID NO: 529; or

(g) a harpin or harpin-like polypeptide and an amino acid sequence ofthe harpin or harpin-like polypeptide comprises any one of SEQ ID NOs:587, 589, 591, 593, 594 and 595; or

(h) a retro inverso harpin or harpin-like polypeptide and an amino acidsequence of the retro inverso harpin or harpin-like polypeptidecomprises any one of SEQ ID NOs: 588, 590, 592, 596 and 597; or

(i) a root hair promoting polypeptide (RHPP) and an amino acid sequenceof the RHPP comprises any one of SEQ ID Nos: 600, 603 and 604; or

(j) a Kunitz Trypsin Inhibitor (KTI) polypeptide and an amino acidsequence of the KTI polypeptide comprises SEQ ID No: 602; or

(k) a retro inverso root hair promoting polypeptide (RI RHPP) and anamino acid sequence of the RI RHPP comprises any one of SEQ ID NO: 601,605 and 606; or

(l) an elongation factor Tu (EF-Tu) polypeptide and an amino acidsequence of the EF-Tu polypeptide comprises any one of SEQ ID NOs:607-623; or

(m) a retro inverso elongation factor Tu (RI EF-Tu) polypeptide and anamino acid sequence of the RI EF-Tu polypeptide comprises any one of SEQID NOs: 624-640; or

(n) a fusion polypeptide comprising SEQ ID NO: 750; or

(o) a phytosulfokine (PSK) polypeptide and an amino acid sequence of thePSK polypeptide comprises SEQ ID NO: 598; or

(p) a retro inverso phytosulfokine (RI PSK) polypeptide and an aminoacid sequence of the RI PSK polypeptide comprises SEQ ID NO: 599; or

(q) a thionin or thionin-like polypeptide and an amino acid sequence ofthe thionin or thionin-like polypeptide comprises any one of SEQ ID NOs:650-749, and

optionally, wherein the flagellin or flagellin-associated polypeptide of(a), the mutant flagellin or flagellin-associated polypeptide of (c),the harpin or harpin-like polypeptide of (g), the PSK polypeptide of(o), and the thionin or thionin-like polypeptide of (q) either: containsa chemical modification; is a variant having an amino acid insertion,deletion, inversion, repeat, duplication, extension, or substitutionwithin the amino acid; is part of a fusion protein; or contains aprotease recognition sequence.

A composition is provided for bioactive priming of a plant or a plantpart to increase growth, yield, health, longevity, productivity, and/orvigor of a plant or a plant part and/or decrease abiotic stress in theplant or the plant part and/or protect the plant or the plant part fromdisease, insects and/or nematodes, and/or increase the innate immuneresponse of the plant or the plant part and/or change plantarchitecture. The composition comprises either: the polypeptide asdescribed above or any combination thereof, and an agrochemical or acarrier; or any combination of the polypeptides.

A seed coated with the polypeptide or the composition as describedherein is also provided.

A recombinant microorganism that expresses or overexpresses apolypeptide is also provided. The polypeptide comprises the polypeptidesas described above for the composition.

Methods are provided for increasing growth, yield, health, longevity,productivity, and/or vigor of a plant or a plant part and/or decreasingabiotic stress in the plant or the plant part and/or protecting theplant or the plant part from disease, insects and/or nematodes, and/orincreasing the innate immune response of the plant or the plant partand/or changing plant architecture. The method can comprise applying thepolypeptide or the composition as described herein to a plant, a plantpart, or a plant growth medium or a rhizosphere in an area surroundingthe plant or the plant part to increase growth, yield, health,longevity, productivity, and/or vigor of the plant or the plant partand/or decrease abiotic stress in the plant or the plant part and/orprotect the plant or the plant part from disease, insects and/ornematodes, and/or increase the innate immune response of the plant orthe plant part and/or change the plant architecture.

Alternatively, the method can comprise applying the polypeptide or thecomposition as described herein to a plant growth medium to increasegrowth, yield, health, longevity, productivity, and/or vigor of a plantor a plant part to be grown in the plant growth medium and/or decreaseabiotic stress in the plant or the plant part to be grown in the plantgrowth medium and/or protect the plant or the plant part to be grown inthe plant growth medium from disease, insects and/or nematodes, and/orincrease the innate immune response and/or change plant architecture ofthe plant or the plant part to be grown in the plant growth medium.

Another method comprises applying the recombinant microorganism asdescribed herein to a plant, a plant part, or a plant growth medium or arhizosphere in an area surrounding the plant or the plant part toincrease growth, yield, health, longevity, productivity, and/or vigor ofthe plant or the plant part and/or decrease abiotic stress in the plantor the plant part and/or protect the plant or the plant part fromdisease, insects and/or nematodes, and/or increase the innate immuneresponse of the plant or the plant part and/or change the plantarchitecture. The recombinant microorganism expresses the polypeptideand expression of the polypeptide is increased as compared to theexpression level the polypeptide in a wild-type microorganism of thesame kind under the same conditions.

A method of producing a polypeptide comprising producing a fusionprotein comprising any polypeptide as described herein and anenterokinase (EK) cleavage site via fermentation, the enterokinasecleavage site enhancing activity and stability of the polypeptide.

The features of the invention are further defined in the appended claimsand the list of embodiments provided below in the Section entitled“EMBODIMENTS.” Other objects and features will be in part apparent andin part pointed out hereinafter.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 shows the Bt.4Q7Flg22 bioactive priming polypeptide in its nativeL configuration (SEQ ID NO: 226) and the corresponding retro inverso orD configuration form (SEQ ID NO: 375).

FIG. 2 illustrates total harvestable yield in corn that received foliarapplications with Bt.4Q7Flg22 (SEQ ID NO:226) in 12 locations (panel A)and retro inverso (RI) version of Bt.4Q7Flg22 (SEQ ID NO: 375) bioactivepriming polypeptides in 10 locations (panel B) and reported in Bu/Ac ascompared to yield in the non-treated control.

FIG. 3 illustrates total harvestable yield in corn that received foliarapplications with Bt.4Q7Flg22 bioactive priming polypeptide (SEQ ID NO:226) in 6 locations and reported in Bu/Ac as compared to yield in thenon-treated control.

FIG. 4 illustrates total harvestable yield in soybean that receivedfoliar applications with Bt.4Q7Flg22 (SEQ ID NO: 226) (panel A) andretro inverso (RI) Bt.4Q7Flg22 (SEQ ID NO: 375) (panel B) bioactivepriming polypeptides in 11 locations and reported in Bu/Ac as comparedto yield in the non-treated control.

FIG. 5 illustrates total harvestable yield in corn that received foliarapplications with Ec.Flg22 (SEQ ID NO: 526) (panel A) and retro inversowith Ec.Flg22 (SEQ ID NO: 527) (panel B) bioactive priming polypeptidesin 12 locations and reported in Bu/Ac as compared to yield in thenon-treated control.

FIG. 6 is directed to a reactive oxygen species (ROS) activity assayusing Bt.4Q7Flg22 in combination with different concentrations ofcellobiose as an additive in corn (panel A) or in soybeans (panel B).

FIG. 7 is directed to a reactive oxygen species (ROS) activity assayusing Bt.4Q7Flg22 at different concentrations to identify the peakactivity and timing for the assay.

FIG. 8 is directed to the application delivery using thionins toinfluence (decrease) the growth of Agrobacterium strain GV3101 in a ratedependent manner.

FIG. 9 is directed to the application delivery of Bt4Q7 Flg22polypeptides tagged or untagged with thionins to decrease the growth ofCandidatus liberibacter spp in HLB infected citrus trees. Data representquantitative PCR results (Ct values) of C. liberibacter in leaf samplestaken from treated infected trees.

FIG. 10 is directed to the application delivery to citrus in treesinjected with 1× or 10× Bt.4Q7Flg22 (SEQ ID NO: 226) to decrease thegrowth of Candidatus liberibacter spp in HLB infected citrus trees. Datarepresent quantitative PCR results (Ct values) of C. liberibacter inleaf samples taken from treated infected trees.

FIG. 11 is directed to ‘Valencia’ orange trees injected with 1× or 10×Bt.4Q7Flg22 (SEQ ID NO: 226) to increase fruit set per limb.

FIG. 12 is directed to ‘Valencia’ orange trees injected with 1× or 10×Bt.4Q7Flg22 (SEQ ID NO: 226) to increase fruit growth as measured incentimeters.

FIG. 13 is directed to ‘Valencia’ orange trees injected with 1× or 10×Bt.4Q7Flg22 (SEQ ID NO: 226) to increase fruit set as indicated byestimated fruit volume per limb.

FIG. 14 is directed to ‘Ruby Red’ grapefruit trees injected with 1× or10× Bt.4Q7Flg22 (SEQ ID NO: 226) to increase fruit set per limb.

FIG. 15 is directed to ‘Ruby Red’ grapefruit trees injected with 1× or10× Bt.4Q7Flg22 (SEQ ID NO: 226) to increase fruit growth as measured incentimeters.

FIG. 16 is directed to ‘Ruby Red’ grapefruit trees injected with 1× or10× Bt.4Q7Flg22 (SEQ ID NO: 226) to increase fruit set as indicated byestimated fruit volume per limb.

DEFINITIONS

When the articles “a,” “an,” “one,” “the,” and “said” are used herein,they mean “at least one” or “one or more” unless otherwise indicated.

The terms “comprising,” “including,” and “having” are intended to beinclusive and mean that there may be additional elements other than thelisted elements.

“Abiotic stress” as used herein is defined as an environmental conditionthat can have a negative impact on a plant. Abiotic stress can include:temperature (high or low) stress, radiation stress (visible or UV),drought stress, cold stress, salt stress, osmotic stress,nutrient-deficient or high metal stress, or water stress that results inwater deficit, flooding or anoxia. Other abiotic stress factors includedehydration, wounding, ozone, and high or low humidity.

“Bioactive priming” refers to an effect of the polypeptides as describedherein to improve a plant or a plant part. Bioactive priming canincrease growth, yield, health, longevity, productivity, and/or vigor ofa plant or a plant part and/or decrease abiotic stress in the plant orthe plant part and/or protect the plant or the plant part from disease,insects and/or nematodes, and/or increase the innate immune response ofthe plant or the plant part and/or change plant architecture.

A “bioactive priming polypeptide” as used herein may be usedinterchangeably with the term “priming agent(s)” and as described forthe classes of polypeptides of the: flagellin and flagellin-associatedpolypeptides, harpin and harpin-like polypeptide (HpaG-like), thionins,elongation factor Tu (EF-Tu) and its polypeptides, phytosulfokine α(PSKα), kunitz trypsin inhibitor (KTI), and root hair promotingpolypeptide (RHPP), as well as any retro inverso polypeptides thereof.

A “colorant” as used herein acts as a visual product identifier forproduct branding and application. Colorants can include, but are notlimited to, dyes and pigments, inorganic pigments, organic pigments,polymeric colorants, and formulated pigment coating dispersionsavailable in a variety of highly concentrated shades.

“Endogenously” applied as used herein refers to an application to theinside of a plant surface. Small bioactive priming polypeptides areparticularly suited for signalling and communication within a plant.Inside a plant surface refers to a surface internal to any plantmembrane or plant cell. Internal could be used to mean eitherextracellular or intracellular to a plant cell and is inclusive ofxylem, phloem, tracheids, etc. Endogenous can refer to movementsystemically or through a plant such as referring to cell to cellmovement in a plant. Endogenous application can include delivery ofbioactive priming polypeptides using recombinant endophytic bacteria orfungi, wherein the endophytic microorganism is delivered externally tothe plant and through natural mechanisms moves internally to the plant.

“Exogenously” applied as used herein refers to an application to theoutside of a plant surface. A plant surface can be any external plantsurface, for example a plasma membrane, a cuticle, a trichome, a leaf, aroot hair, seed coat, etc.

“-associated” or “-like” polypeptides as used herein refers topolypeptides derived from or structurally similar to the recitedpolypeptide but having an amino acid sequence and/or source distinctfrom the recited polypeptide. For example, the thionin-like protein fromBrassica rapa (SEQ ID NO: 694) has a different sequence than thioninfrom Brassica napus (SEQ ID NOs 693) but is structurally andfunctionally similar.

A “foliar treatment” as used herein refers to a composition that isapplied to the above ground parts or foliage of a plant or plant partand may have leaves, stems, flowers, branches, or any aerial plant part,for example, scion.

“Injection” as described herein can be used interchangeably withvaccination or immunization and provides a process whereby the bioactivepriming polypeptides are delivered endogenously to a plant or plantpart.

“Inoculation” means to deliver-bacteria or living microorganisms thatproduce the priming polypeptide to a plant or plant part. Inoculationcan also refer to the delivery of the priming polypeptide for passiveentry through the stomata or any opening in or on a plant or plantpart.A “plant” refers to but is not limited to a monocot plant, a dicotplant, or a gymnosperm plant. The term “plant” as used herein includeswhole plants, plant organs, progeny of whole plants or plant organs,embryos, somatic embryos, embryo-like structures, protocorms,protocorm-like bodies, and suspensions of plant cells. Plant organscomprise, shoot vegetative organs/structures (e.g., leaves, stems andtubers), roots, flowers and floral organs/structures (e.g., bracts,sepals, petals, stamens, carpels, anthers and ovules), seed includingembryo, endosperm, and seed coat and fruit (the mature ovary), planttissue (e.g., phloem tissue, vascular tissue, ground tissue, and thelike) and cells (e.g., guard cells, egg cells, trichomes and the like).The class of plants that can be used in the methods described herein isgenerally as broad as the class of higher plants, specificallyangio-sperms monocotyledonous (monocots) and dicotyledonous (dicots)plants and gymnosperms. It includes plants of a variety of ploidylevels, including aneuploid, polyploid, diploid, haploid, homozygous andhemizygous. The plants described herein can be monocot crops, such as,sorghum, maize, wheat, rice, barley, oats, rye, millet, and triticale.The plants described herein can also be dicot crops, such as apple,pear, peach, plum, orange, lemon, lime, grapefruit, kiwi, pomegranate,olive, peanut, tobacco, tomato, etc. Also, the plants can behorticultural plants such as rose, marigold, primrose, dogwood, pansy,geranium, etc.

A plant “biostimulant” is any substance or microorganism applied to aplant or a plant part that is used to enhance nutrition efficiency,abiotic stress tolerance and/or any other plant quality trait(s).

A “plant cell” as used herein refers to any plant cell and can comprisea cell at the plant surface or internal to the plant plasma membrane,for example, an epidermal cell, a trichome cell, a xylem cell, a phloemcell, a sieve tube element, or a companion cell.

A “plant part” as described herein refers to a plant cell, a leaf, astem, a flower, a floral organ, a fruit, pollen, a vegetable, a tuber, acorm, a bulb, a pseudobulb, a pod, a root, a rhizome, a root ball, aroot stock, a scion, or a seed.

A “polypeptide” as described herein refers to any protein, peptide orpolypeptide.

“Priming” or “peptide priming” as used herein refers to a technique usedto improve plant performance. In particular priming is a process wherebythe bioactive priming polypeptides are applied either exogenously orendogenously to a plant, plant part, plant cell or to the intercellularspace of a plant that results in outcomes that provide benefits to aplant, such as enhanced growth, productivity, abiotic stress tolerance,pest and disease tolerance or prevention.

A “retro-inverso” polypeptide as used herein refers to a polypeptidechain of a natural derived polypeptide from a normal-all-L chainreconfigured and built using non-naturally occurring D-amino acids inreverse order of the naturally occurring L-amino acids. The all-D-aminoacid form and the parent chain containing all L-form are topologicalmirrorings of the protein structure.

A “seed treatment” as used herein refers to a substance or compositionthat is used to treat or coat a seed. Sample seed treatments include anapplication of biological organisms, chemical ingredients, inoculants,herbicide safeners, micronutrients, plant growth regulators, seedcoatings, etc, provided to a seed to suppress, control or repel plantpathogens, insects, or other pests that attack seeds, seedlings orplants or any useful agent to promote plant growth and health.

A “synergistic” effect refers to an effect arising between theinteraction or cooperation of two or more bioactive primingpolypeptides, substances, compounds, or other agents to produce acombined effect greater than the sum of their separate effects.

A “synergistic effective concentration” refers to the concentration(s)of two or more bioactive priming polypeptides, substances, compounds orother agents that produces an effect greater than the sum of theindividual effects.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

There is a growing need for bioactive polypeptides that act as “primingagents” to provide benefits to agriculture. The use of bioactive“priming” polypeptides in agricultural practices provides a paradigmshift for integrated crop management practices for example, to managedisease, abiotic stress and yield programs. Bioactive (naturallyoccurring, recombinant or synthetic) priming polypeptides are deliveredin agricultural formulations. Compositions and methods of using thebioactive priming polypeptides are described to supply a multi-tieredtreatment regime to apply to crops to achieve agronomically desirableoutcomes. Such desirable outcomes include enhanced phenotypes in plantssuch as those that exhibit protection against pest, disease agents andabiotic stress, as well as increased plant growth, productivity andyield. More specifically, the bioactive priming polypeptides orformulations of the bioactive priming polypeptides can be applied usingvarious treatment regimes, exogenously and/or endogenously to a plant orplant part, and have been discovered to increase growth, yield, health,longevity, productivity, and/or vigor of a plant or a plant part and/ordecrease abiotic stress in the plant or the plant part and/or protectthe plant or the plant part from disease, insects and/or nematodes,and/or increase the innate immune response of the plant or the plantpart and/or change plant architecture.

Specific classes of synthetically derived or naturally occurringbioactive priming polypeptides including flagellins andflagellin-associated polypeptides (including those conserved among theBacillus genera), thionins, harpin-like polypeptide (HpaG-like),elongation factor Tu (EF-Tu), phytosulfokine (PSKα) and root hairpromoting polypeptide (RHPP) were selected for their distinct modes ofaction and can be used individually or in combination with otherpolypeptides to accommodate the specific agricultural needs describedabove. They can be used in the place of or in addition to commerciallyavailable agrochemicals, biostimulants, supplemental bioactives and/orpesticidal compounds.

Combinations of the bioactive priming polypeptides are also providedthat are applied in synergistically effective amounts to provide controlof pests, pathogens and additionally provide benefits to enhance plantgrowth and promote plant health.

I. Polypeptides

The bioactive priming polypeptides are provided as naturally occurring,recombinant or chemically synthesized forms derived from bacteria orplants. The bioactive priming polypeptides are provided in both thenormal L and non-natural retro-inverso D amino-acid forms. In addition,bioactive priming polypeptides are provided that contain non-naturalmodifications, including N-terminal and C-terminal modifications,cyclization, β-amino and D-amino acid containing, and other chemicalmodifications that enhance stability or performance of the polypeptides.For example, flagellin and the Flg-associated polypeptides comprising 22amino acids in length and derived from the full coding region offlagellin were initially isolated and identified from a proprietarygenome assembled for bacterial strain, Bacillus thuringiensis 4Q7. TheseFlg22 derived polypeptides were provided in the standard (L) andretro-inverso (D) forms. They are described as Bt.4Q7Flg22 andretro-inverso (RI) Bt.4Q7Flg22. Other bacterial derived bioactivepriming polypeptides are Ec.Flg22 (Escherichia coli), HpaG-like(Xanthomonas spp.), while the plant derived polypeptides includethionins (Citrus spp. and other plant species), PSKa (Arabidopsisthaliana and other plants), EF-Tu (both bacterial or plant derived) andRHPP (Glycine max).

The bioactive priming polypeptides can include full-length proteins andare provided as naturally occurring, synthetic or recombinant formsderived from bacteria or plants. For example, flagellin, EF-Tu, KTI, andHpaG can all be delivered to plants.

The bioactive priming polypeptides can also be delivered as fusionpartners to other protein sequences, including protease cleavage sites,binding proteins, and targeting proteins for specific delivery to plantsor plant parts.

Also provided are signature, signal anchor sorting and secretionsequences that can be naturally or chemically synthesized and targetingsequences, such as phloem-targeting sequences that are produced alongwith the bioactive priming polypeptide(s) using recombinantmicroorganisms and either used as fusion or assistance polypeptides withthe bioactive priming polypeptides as described herein.

Non-naturally occurring polypeptides are also described herein. Morespecifically, a polypeptide is provided for bioactive priming of a plantor a plant part to increase growth, yield, health, longevity,productivity, and/or vigor of a plant or a plant part and/or decreaseabiotic stress in the plant or the plant part and/or protect the plantor the plant part from disease, insects and/or nematodes, and/orincrease the innate immune response of the plant or the plant partand/or change plant architecture. The polypeptide comprises either:

(a) a flagellin or flagellin-associated polypeptide and an amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 1-225, 227-375, 526, 528, 530, 532, 534,536, 538, 540, and 541; or

(b) a mutant flagellin or flagellin-associated polypeptide and an aminoacid sequence of the mutant flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 571-579; or

(c) a mutant flagellin or flagellin-associated polypeptide and an aminoacid sequence of the mutant flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 580-586; or

(d) a retro inverso Flg22 polypeptide and an amino acid sequence of theretro inverso Flg22 polypeptide comprises any one of SEQ ID NOs:376-450, 527, 531, 533, 535, 537 and 539; or

(e) a retro inverso FlgII-28 polypeptide and an amino acid sequence ofthe retro inverso FlgII-28 polypeptide comprises any one of SEQ ID NOs:451-525; or

(f) a retro inverso Flg15 polypeptide and an amino acid sequence of theretro inverso Flg15 polypeptide comprises SEQ ID NO: 529; or

(g) a harpin or harpin-like polypeptide and an amino acid sequence ofthe harpin or harpin-like polypeptide comprises any one of SEQ ID NOs:587, 589, 591, 593, 594 and 595; or

(h) a retro inverso harpin or harpin-like polypeptide and an amino acidsequence of the retro inverso harpin or harpin-like polypeptidecomprises any one of SEQ ID NOs: 588, 590, 592, 596 and 597; or

(i) a root hair promoting polypeptide (RHPP) and an amino acid sequenceof the RHPP comprises any one of SEQ ID Nos: 600, 603 and 604; or

(j) a Kunitz Trypsin Inhibitor (KTI) polypeptide and an amino acidsequence of the KTI polypeptide comprises SEQ ID No: 602; or

(k) a retro inverso root hair promoting polypeptide (RI RHPP) and anamino acid sequence of the RI RHPP comprises any one of SEQ ID NO: 601,605 and 606; or

(l) an elongation factor Tu (EF-Tu) polypeptide and an amino acidsequence of the EF-Tu polypeptide comprises any one of SEQ ID NOs:607-623; or

(m) a retro inverso elongation factor Tu (RI EF-Tu) polypeptide and anamino acid sequence of the RI EF-Tu polypeptide comprises any one of SEQID NOs: 624-640; or

(n) a fusion polypeptide comprising SEQ ID NO: 750; or

(o) a phytosulfokine (PSK) polypeptide and an amino acid sequence of thePSK polypeptide comprises SEQ ID NO: 598; or

(p) a retro inverso phytosulfokine (RI PSK) polypeptide and an aminoacid sequence of the RI PSK polypeptide comprises SEQ ID NO: 599; or

(q) a thionin or thionin-like polypeptide and an amino acid sequence ofthe thionin or thionin-like polypeptide comprises any one of SEQ ID NOs:650-749, and

optionally, wherein the flagellin or flagellin-associated polypeptide of(a), the mutant flagellin or flagellin-associated polypeptide of (c),the harpin or harpin-like polypeptide of (g), the PSK polypeptide of(o), and the thionin or thionin-like polypeptide of (q) either: containsa chemical modification; is a variant having an amino acid insertion,deletion, inversion, repeat, duplication, extension, or substitutionwithin the amino acid; is part of a fusion protein; or contains aprotease recognition sequence.

Flagellins and Flagellin-Associated Polypeptides

The polypeptide can include a flagellin or flagellin-associatedpolypeptide.

The flagellin or flagellin-associated polypeptide can be derived from aBacillus, a Lysinibacillus, a Paenibacillus, an Aneurinibacillus genusbacterium, or any combination thereof.

One of the main classes of bioactive priming polypeptides as describedherein are the flagellin(s) and the flagellin-associated primingpolypeptide(s). Conserved full and partial length amino acid flagellincoding sequences were identified from various species of Bacillus andnon-Bacillus bacteria using methods as described herein.

Flagellin is a structural protein that forms the main portion offlagellar filaments from flagellated bacterial species that can showconservation in the N-terminal and C-terminal regions of the protein butcan be variable in the central or mid part (Felix G. et al., “Plantshave a sensitive perception system for the most conserved domain ofbacterial flagellin,” The Plant Journal 18: 265-276, 1999). The N- andC-terminal conserved regions from flagellins that form the inner core ofthe flagellin protein may have roles in the polymerization of theprotein into a filament, in the motility and transport of the proteinand in the surface attachment of a peptide fragment to the plant cellmembrane/cell surface receptors of a plant.

Full or partial flagellins (Table 1-2) and the flagellin-associatedpolypeptides derived from those Bacillus and non-Bacillus flagellins(Tables 3 and 5) are provided.

The amino acid sequence of the flagellin or flagellin-associatedpolypeptide can comprise any one of SEQ ID NOs: 1-768, or anycombination thereof.

Flagellin-associated bioactive priming polypeptides are produced fromflagellin coding polypeptides (such as the precursor proteins of Flg22).More specifically, a polypeptide or a cleaved fragment derived from thepolypeptide is provided to achieve a bioactive priming Flg polypeptidethat can be used to prime or treat a plant. The cleavage of the Flg22fragment from larger precursors can be accomplished through introductionof proteolytic cleavage sites near the Flg22 to facilitate processing ofthe active biopeptide from the larger polypeptide.

The flagellin-associated bioactive priming polypeptides can be derivedfrom full length flagellin proteins (or precursor proteins fromFlg-associated polypeptides from a Bacillus, a Lysinibacillus, aPaenibacillus, or an Aneurinibacillus or other non-related generabacterium). For example, PCR purified DNA from the flagellin-associatedpolypeptides such as Flg22 and FlgII-28 (Bacillus genera) and Flg15 andFlg22 (E. coli) are cloned into a recombinant vector, amplified toachieve adequate amounts of purified DNA that is then sequenced usingconventional methods known and used by one of ordinary skill in the art.The same methods can be used with the flagellin coding or the flagellinpartial sequences (Table 1), N- or C-terminal flagellin polypeptides(Table 2) and any of the Flg-associated polypeptides (Tables 3-5).

The flagellin or flagellin-associated polypeptide can be derived fromany member of Eubacteria that contains the conserved 22 amino acidregion that is recognized by the plants. Preferred flagellin orflagellin-associated polypeptides can be derived from a Bacillus, aLysinibacillus, a Paenibacillus, an Aneurinibacillus genus bacterium, orany combination thereof. Additional preferred flagellin and Flg22sequences can be obtained from the gammaproteobacteria, which containconserved 22 amino acid sequences of >68% identity.

Conserved Flagellin Sequences from Bacillus

The flagellin-associated bioactive priming polypeptides correspond tothe N-terminal conserved domains of Bacillus spp. and other Eubacterialflagellin and are provided as synthetic, recombinant or naturallyoccurring forms. The flagellin bioactive priming polypeptides of Flg22,Flg15 and FlgII-28 (Table 3) were identified and act as potent elicitorson a wide range of crops and vegetables to prevent and treat the spreadof select disease(s) while synergistically stimulating and promotinggrowth responses in plants.

The flagellin and flagellin-associated bioactive priming polypeptides asdescribed herein are provided for use individually or in combinationwith other bioactive priming polypeptides as described herein, andinclude conserved full and partial flagellins from Bacillus (Table 1),conserved N- and C-terminal regions from flagellin polypeptides (Table2), Bacillus derived Flg22 and FlgII-28-derived bioactive primingpolypeptides (Table 3) and retro-inverso sequences that are mirrorimages derived from the Bacillus Flg22 and FlgII-28 (Table 4). Theunderlined portion of the sequences in Tables 1 and 3 representidentified signal anchor sorting or secretion sequences, and signalanchoring sequences, respectively. Other non-Bacillus derivedpolypeptide and proteins are also described that are functionalequivalents and can be utilized in similar fashion (Table 5).

TABLE 1 Conserved flagellin sequences from Bacillus SEQ ID NO:Full or Partial Flagellin Coding Sequence-Amino Acid FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINSASDDAAGLAIATRM SEQ ID NO: 1KAREGGLNVAGRNTQDGMSLIRTADSALNSVSNILLRMRDLANQSANGTNTK BacillusGNQASLQKEFAQLTEQIDYIAKNTQFNDQQLLGTADKKIKIQTLDTGSTNPA thuringiensisQIEITLNSVKSADLGLDVQIGDEGDAESTAAADPTSAKQAIDAIDAAITTVA strain 4Q7GQRATLGATLNRFEFNANNLKSQETSMADAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINSASDDAAGLAIATRM SEQ ID NO: 2KAREGGLNVAGRNTQDGMSLIRTADSALNSVSNILLRMRDLANQSANGTNTK BacillusGNQASLQKEFAQLTEQIDYIAKNTQFNDQQLLGTADKKIKIQTLDTGSTNPA thuringiensis,QIEITLNSVKSADLGLDVQIGDEGDAESTAAADPTSAKQAIDAIDAAITTVA strain HD1002GQRATLGATLNRFEFNANNLKSQETSMADAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINSASDDAAGLAIATRM SEQ ID NO: 3KAREGGLNVAGRNTQDGMSLIRTADSALNSVSNILLRMRDLANQSANGTNTK BacillusGNQASLQKEFAQLTEQIDYIAKNTQFNDQQLLGTADKKIKIQTLDTGSTNPA thuringiensis,QIEITLNSVKSADLGLDVQIGDEGDAESTAAADPTSAKQAIDAIDAAITTVA strain HD-789GQRATLGATLNRFEFNANNLKSQETSMADAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINSASDDAAGLAIATRM SEQ ID NO: 4KAREGGLNVAGRNTQDGMSLIRTADSALNSVSNILLRMRDLANQSANGTNTK Bacillus cereusGNQASLQKEFAQLTEQIDYIAKNTQFNDQQLLGTADKKIKIQTLDTGSTNPA strain G9842QIEITLNSVKSADLGLDVQIGDEGDAESTAAADPTSAKQAIDAIDAAITTVAGQRATLGATLNRFEFNANNLKSQETSMADAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRIGTNVLSMNARQSLYENEKHMNVAMEHLATGKKLNNASDNPANIAIVTRM SEQ ID NO: 5HARASGMRVAIRNNEDAISMLRTAEAALQTVTNILQRMRDLAVQSANGTNSN Bacillus KNRHSLNKEFQSLTEKIGYIGETTEFNDLSVFEGQNRPITLDDIGHTINMMK thuringiensisHIPPSPTQHDIKISTEQEARAAILKIEDALQSVSLHRADLGAMINRLQFNIE serovarindianaNLNSQSMALTDAASLIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQMV strain HD521 SKLLQSFlagellin MRINTNINSMRTQEYMRQNQAKMSNSMDRLSSGKRINNASDDAAGLAIATRMSEQ ID NO: 6 RARESGLGVAADNTQNGMSLIRTADSAMNSVSNILLRMRDIANQSANGTNTNBacillus ENKSALQKEFAQLQKQITYIAENTQFNDKNLLNEDSEVKIQTLDSSKGEQQIthuringiensis TIDLKAVTLEKLNIKDIAIGKADAADKPVTPGATVDQKDLDSVTDKIAALTEstrain CTC TSSKADIDAIQSSLDNFKASMTPEDVKTLEDALKGFKTGQANPADAGVDAIQDALSKVKLPTATAAAPAADADKSDALAAIAAIDAALTKVADNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQAN QTPQMVSKLLQFlagellin MTGITINLEIDFFAYYRFSICRKVNIKKWGFLNMRINTNINSMRTQEYMRQNSEQ ID NO: 7 QAKMSNAMDRLSSGKRINNASDDAAGLAIATRMRARENGLGVAANNTQDGMSBacillus LIRTADSAMNSVSNILLRMRDLANQSANGTNTDDNQKALDKEFSALKEQIDYthuringiensis ISKNTEFNDKKLLNGENKTIAIQTLDNADTTKQININLADSSTSALQIDKLTserovaryunnanensis ISGKTTDTTKTETITVTDDEIKAAKTDIDEFNDAKKALADLKAETSAGKADGstrain IEBC-T20001 STDDEIKTAVSNFTKSFEKIQKFMNDSDIKTVQTEIEKFDAAAPALDKAKGMGIAFTSAMDPKAGTITKAATRQNASDAIKSIDAALETIASNRATLGATLNRLDFNVNNLKSQSSSMAAAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANV FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 8RARENGLGVAANNTQDGMSLIRTADSALQSVSNILLRMRDLANQSANGTNTD Bacillus ENKAAMEKEFGQLKDQIKYITDNTQFNDKNLLDAASGTTKSIAIQTLDSDQA thuringiensisSTQIEIKIAGSSLAALGLDKVQIGQETVAQKDLDVLTKAMGRLAAPDADATT serovar tolworthiRDLDVQVAKDAFDKVKGFIADPAQAKAVERAFEDYTAAEAGKEEDAAKAIDAAYKKVTGLTAGTTGTVDAHNAVNKIDAALKTVADNRATLGATLNRLDFNVNNLKSQSASMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVS KLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNSMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 9RARESGLGVAANNTQDGMSLIRTADSAMNSVSNILLRMRDIANQSANGTNTD Bacillus cereus KNQVALQKEFGELQKQIDYIAKNTQFNDKNLLSGKAGAPDQALEINIQTLDS strain FM1SDPNQQIKISLDSVSTAQLGVKDLQIGSSSITQQQLDTLDNAMKRLETASTTAAVRDQDVADAKAAFENVKGFFSEGNVDSINRAFTDFANETTNKDDKAEAIYALYNNATLITKPTPDASNPASVDPANAIKKIDQAIEKIASSRATLGATLNRLDFNVNNLKSQQSSMASAASQVEDADMAKEMSEMTKFKILNEAGISMLSQANQ TPQMVSKLLQFlagellin MRIGTNVLSMNARQSFYENEKRMNVAIEHLATGKKLNHASDNPANVAIVTRMSEQ ID NO: 10 HARTSGIHVAIRNNEDAISMLRTAEAALQTVTNILQRMRDVAVQSANGTNSNBacillus cereus KNRDSLNKEFQSLTEQIGYIDETTEFNDLSVFDRQNCPVTLDDIGHTVNVTKstrain FM1 HIPPSPTQHDINISTEQEARAAIRKIEETLQNVSLHRADLGAMINQLQFNIENLNSQSTALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQMV YKLLQS FlagellinMDRLSSGKRINNASDDAAGLAIATRMRARESGLGVAANNTQDGMSLIRTADS SEQ ID NO: 11ALNSVSNILLRMRDIANQSANGTNTADNQQALQKEFGQLKEQISYIADNTEF BacillusNDKTLLKADNSVKIQTLDSADTNKQISIDLKGVTLNQLGLDTVNIGSEKLSA thuringiensisESLNVAKATMARLVKADQNADPSTFALDVNTAKESFDKIKGFIANKTNVQNV strain MC28ENAFNDYAVADPADKADKADAIQAAFNTAITGLTAGTPNTSNPSSAVDSIDAALKTVASNRATLGATLNRLDFNVNNLKSQSASMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNSMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 12RSREGGLNVAARNTEDGMSLIRTADSALNSVSNILLRMRDLANQSASGTNTD BacillusKNQAAMQKEFDQLKEQIQYIADNTEFNDKKLLDGSNSTINIQTLDSHDKNKQ bombysepticusITISLDSASLKNLDIKDLAIGSATINQTDLDTATNSMKRLATPATDGKVLAQ strain WangDIADAKAAFNKVQSAYTPAEVDKIQDAFKAYDKLAADPASKATDIADAAKNVNTVFGTLATPTATKFDPSSAVEKIDKAIETIASSRATLGATLNRLDFNVTNLKSQENSMAASASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSK LLQ FlagellinMTGITINLEIDFFAYYRFSICRKVNIKKWGFLNMRINTNINSMRTQEYMRQN SEQ ID NO: 13QAKMSNSMDRLSSGKRINNASDDAAGLAIATRMRSREGGLNVAARNTEDGMS BacillusLIRTADSALNSVSNILLRMRDLANQSASGTNTDKNQAAMQKEFDQLKEQIQY thuringiensisIADNTEFNDKKLLDGSNSTINIQTLDSHDKNKQITISLDSASLKNLDIKDLA serovar kenyaeIGSATINQTDLDTATNSMKRLATPATDGKVLAQDIADAKAAFNKVQSAYTPAEVDKIQDAFKAYDKLAADPASKDTDIADAAKNVNTVFGTLATPTATKFDPSSAVEKIDKAIETIASSRATLGATLNRLDFNVTNLKSQENSMAASASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNSMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 14RSREGGLNVAARNTEDGMSLIRTADSALNSVSNILLRMRDLANQSASGTNTD BacillusKNQAAMQKEFDQLKEQIQYIADNTEFNDKKLLDGSNSTINIQALDSHDKNKQ thuringiensisITISLDSASLKNLDIKDLAIGSATINQTDLDTATNSMKRLATPATDGKVLAQ serovar kenyaeDIADAKAAFNKVQSAYTPAEVDKIQDAFKAYDKLAADPASKDTDIADAAKNVNTVFGTLATPTATKFDPSSAVEKIDKAIETIASSRATLGATLNRLDFNVTNLKSQENSMAASASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSK LLQFlagellin (A-type) MRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRMSEQ ID NO: 15 RARENGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDLANQSANGTNTGBacillus cereus DNQKALDKEFSALKEQIDYISKNTEFNDKKLLNGDNKTIAIQTLDNADTSKQININLADSSTSALKIEKLTISGSTAIAGKTEKVTITAEDIKAAEEDIKAFTQAQEGLANLVKEVKDTDGSVKTPGSTPDDIKKAVTAFTESFEKMKKFMNDEDITKVEEKIKAFDAASPDLDAAKEMGTAFTAAMKPAAGEITKAAMKPNASDAIKSIDEALETIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ Flagellin (A-type)MRIGTNVLSMNARQSLYENEKRMNVAMEHLATGKKLNNASDNPANIAIVTRM SEQ ID NO: 16HARASGMRLAIRNNEDTISMLRTAEAALQTLTNILQRMRDLAVQSANGTNSN Bacillus cereusKNRDSLNKEFQSLTEQIGYIGETTEFNDLSVFDGQNRPVTLDDIDHTINMTKHIPPSPTQHDIKISTEQEARAAILKIEEALQSVSIHRADLGSMINRLQFNIENLNSQSMALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQMV SKLLQS FlagellinMRIGTNVLSMNARQSLYENEKRMNVAMEHLATGKKLNHASDNPANVAIVTRM SEQ ID NO: 17HARASGMRVAIRNNEDAISMLRTAEAALQTVTNVLQRMRDVAVQSANGTNLN BacillusKNRDSLNNEFQSLTEQIGYIDETTAFNDLSVFDGQNRPVTLDDIGHTVNVTK thuringiensisHISPSPTQHDINISTEQEARAAIRKIEEALQNVSLYRADLGAMINRLQFNIE serovar finitimusNLNSQSTALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQMV strain YBT-020YKLLQS Flagellin MRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRMSEQ ID NO: 18 RARESGLNVAADNTQNGMSLIRTADSAMNSVSNILLRMRDIANQSANGTNTDBacillus SNKSALQKEFAELQKQITYIADNTQFNDKNLLKEDSEVKIQTLDSSKGEQQIthuringiensis GIDLKAVTLEKLGINNISIGKADGTTEGTKADLTALQAAAKKLEKPDTGTMEserovar finitimus KDVKDAKEEFDKVKASLSDEDVKKIEAAFGEFDKDKTNTTKASDIFNAIKDVstrain YBT-020 KLADKAAAAPAPADLTKFKAALDKLQTPNAGTMVDDVKDAKDEFEKIKGSLSDADAQKIQAAFEEFEKANTDDSKASAIYNLAKDVKVNATDTTTGTDKDTTTSTDKDAALAAIAAIDAALTKVADNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 19RARESGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDLANQSANGTNTA Bacillus cereusENKAAMQKEFGELKDQIKYISENTQFNDQHLLNAAKGSTNEIAIQTLDSDSS stain B4264SKQIKITLQGASLDSLDIKDLQIGSGSTVSQTDLDVLDATMTRVKTATGATRDVDVQAAKSAFDKVKGLMTKPAEVKAIERAFEDYNAGKTDALATAIEAAYTANKTGLPAPAAAAGTVDALGAITKIDAALKTVADNRATLGATLNRLDFNVNNLKSQSASMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSK LLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 20RARESGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDIANQSANGTNTS BacillusDNQKALDKEFSALKEQIDYISKNTEFNDKKLLNGDNKSIAIQTLDNADTTKQ thuringiensisININLADSSTTALNIDKLSIEGTGNKTITLTAADIAKDKANIDAVGTAKTAL serovarAGLTGTPAAAAINSAVADFKTAFAKADKNLMSDAQIKAVTDAITAFEADATP nigeriensisDLTKAKAIGTAYTAPAAGDITKASPNASEAIKSIDAALDTIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQ ANQTPQMVSKLLQFlagellin MRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRMSEQ ID NO: 21 RARESGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDIANQSANGTNTABacillus  DNQQALQKEFGQLKEQISYIADNTEFNDKTLLKADNSVKIQTLDSADTNKQIthuringiensis SIDLKGVTLNQLGLDTVNIGSETLSAESLNVAKATMARLVKADQNADPSTFALDVNTAKESFDKIKGFITNKTNVQNVENAFNDYTVADPADKADKADAIQAAFNTAITGLTAGTPNTSNPSSAVDAIDAALKTVASNRATLGATLNRLDFNVNNLKSQSASMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSK LLQ FlagellinMRIGTNVLSMNARQSLYENEKRMNVAMEHFATGKKLNHASDNPANVAIVTRM SEQ ID NO: 22HARASGMRVAIRNNEDAISMLRTAEAALQTVMNILQRMRDLAVQSANGTNSN BacillusKNRDSLNKEFQSLTEQIGYIGETTEFNDLSVFDGQNRPVTLDDIGHTVNVTK thuringiensisHTSPSPTKHDIKISTEQEARAAIRKIEEALQNVSLHRADFGAMINRLQFNIE serovar konkukianNLNSQSMALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQMV strain 97-27 SKLLQSFlagellin MRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRMSEQ ID NO: 23 RARESGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDIANQSANGTNTABacillus DNQQALQKEFGQLKEQISYIADNTEFNDKTLLKADNSVKIQTLDSADTNKQIthuringiensis SIDLKGVTLNQLGLDTVNIGSETLSAESLNVAKATMARLVKADQNADPSTFAserovar LDVNTAKESFDKIKGFITNKTNVQNVENAFNDYTVADPADKADKADAIQAAF konkukianNTAITGLTAGTPNTSNPSSAVDAIDAALKTVASNRATLGATLNRLDFNVNNL strain 97-27KSQSASMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSK LLQFlagellin protein MRIGTNVLSMNARQSLYENEKRMNVAMEHLATGKKLNHASDNPANIVIVTRMFlaA YARASGMRVAIRNNEDAISMLRTAEAALQTVTNILQHMRDFAIQSANGTNSN SEQ ID NO: 24TNRDSLNKEFQSLTEPIGYIGETTEFNDLSVFDGQNRPITLDDIGHTINMTK Bacillus HIPPSPTQHDIKISTEQEARAAIRKIEEALQNVSLHRADLGSMINRLQFNIE thuringiensisNLNSQSMALIDTASQVEDADMAQEISDFLKFKLLTAVALSVVSQANQIPQIV serovar  SKLLQSthuringiensis strain IS5056 Flagellin proteinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM FlaARARESGLGVAANNTQDGMSLIRTADSAMNSVSNILLRMRDISNQSANGTNTD SEQ ID NO: 25KNQSALDKEFAALKDQIDYISKNTEFNDQKLLDGSKKSIAIQTLDNADTNKQ BacillusIDIQLSNVSTKELKLDTLSIEGSSSKTFTITADDMLAVGTANATAKAKAGTL thuringiensisKGLNVTTGDLTAAKTDVQDFRAAFDKVKGFMGSTEVTNIEKALTKFDGDQSL serovar ANAKAIGDALTSDLATTIAKDQTYSKNVSNASSAIASIDAALESIASNRATL thuringiensisGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGIS strain IS5056MLSQANQTPQMVSKLLQ Flagellin BMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 26RARESGLGVAANNTQDGMSLIRTADSAMNSVSNILLRMRDISNQSANGTNTD BacillusKNQSALDKEFAALKDQIDYISKNTEFNDQKLLDGSKKSIAIQTLDNADTNKQ thuringiensisIDIQLSNVSTKELKLDTLSIEGSSSKTFTITADDMLAVGTANATAKAKAGTL strain Bt407KGLNVTTGDLTAAKTDVQDFRAAFDKVKGFMGSTEVTNIEKALTKFDGDQSLANAKAIGDALTSDLATTIAKDQTYSKNVSNASSAIASIDAALESIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGIS MLSQANQTPQMVSKLLQFlagellin MRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRMSEQ ID NO: 27 RARESGLGVAANNTQDGMSLIRTADSAMNSVSNILLRMRDISNQSANGTNTDBacillus  KNQSALDKEFAALKDQIDYISKNTEFNDQKLLDGSKKSIAIQTLDNADTNKQthuringiensis IDIQLSNVSTKELKLDTLSIEGSSSKTFTITADDMLAVGTANATAKAKAGTLserovar chinensis KGLNVTTGDLTAAKTDVQDFRAAFDKVKGFMGSTEVTNIEKALTKFDGDQSLCT-43 ANAKAIGDALTSDLATTIAKDQTYSKNVSNASSAIASIDAALESIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGIS MLSQANQTPQMVSKLLQFlagellin MTGITINLEIDFFAYYRFSICRKVNIKKWGFLNMRINTNINSMRTQEYMRQNSEQ ID NO: 28 QAKMSNAMDRLSSGKRINNASDDAAGLAIATRMRARESGLGVAANNTQDGISBacillus LIRTADSAMNSVSNILLRMRDLANQSANGTNTNENQAALNKEFDALKEQIDYthuringiensis ISTNTEFNDKKLLDGSNKTIAVQTLDNADTSKQININLSNVSTKELGLDTLSserovar IGTDKVEKTVYDATTKAFADLGAKTGADKAAFDADVTAAMKEFDKVKPFMSA canadensisDDVKKIETKLEDYNKANDAGAQTAAQALGKEFATLTKLETTDLKANASGAIASIDTALKNIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMTGITINLEIDFFAYYRFSICRKVNIKKWGFLNMRINTNINSMRTQEYMRQN SEQ ID NO: 29QAKMSNAMDRLSSGKRINNASDDAAGLAIATRMRARESGLGVAANNTQDGIS BacillusLIRTADSAMNSVSNILLRMRDLANQSANGTNTNENQAALNKEFDALKEQIDY thuringiensisISTNTEFNDKKLLDGSNKTIAVQTLDNADTSKQININLSNVSTKELGLSTLS serovarIGTDKVEKTVYDATTKAFADLGAKTGTDKAAFAADVTAAMKEFDKVKPFMSA galleriaeDDVKKIETKLEDYNKANDAGAEAAAQALGKEFATLTKLETTDLKANASGAIASIDTALKNIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM N-terminalRARESGLSVAANNTQDGMSLIRTADSAMNSVSNILLRMRDLSNQSANGTNTD helical regionENQQALNKEFAALKDQIDYISKNTEFNDKKLLDGSNKSIAIQTLDNADTTKQ SEQ ID NO: 30INIDLSNVSTDTLNISGLTINGKKDITVTISDKDIANAATDIGKATSAQQGL BacillusADLTDTTPAVPDTPAVIGTGTAGNPQFPAVKGTPEIPGSSPAEIAKAVDDFK weihenstephanensisQAFNKVKGLMSDSAVSAMEQKFATFEKDKSLANAKDIGTAFSAPIAGNITKGEQNASGAIKSIDAALEKIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMTGITINLEIDFFAYYRFSICRKVNIKKWGFLNMRINTNINSMRTQEYMRQN SEQ ID NO: 31QAKMSNAMDRLSSGKRINNASDDAAGLAIATRMRARESGLGVAANNTQDGMS BacillusLIRTADSALNSVSNILLRMRDIANQSANGTNTGDNQKALDKEFSALKEQIDY thuringiensisISKNTEFNDKKLLNGDNKSIAIQTLDNADTAKQININLADSSTKALNIDTLS serovar ostriniaeIAGTTDKTITITAKDLTDNKTTLDALKTAKDDLAKLDDKSDQATIDKAVDAFKTAFNNVDKNLLSDKAIEGITEKMTAFDGTHTAAAAIGAAYTEPTAADIKKSAPNASGAIKSIDAALETIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRIGTNVLSMNARQSLYENEKRMNVAMEHLATGKKLNHASDNPANVAIVTRM SEQ ID NO: 32HARASGMRVAIRNNEDALSMLRTAEATLQTVANILQRMRDLAVQSSNDTNSN BacillusKNRDSLNKEFQSLTEQISYIGETTEFNDLSVFDGQNRPVTLDDIGHTVNVTK thuringiensisHISPSPTQHDIKISTEQEARAAIRKIEEALQNVLLHRADLGAMINRLQFNIENLNSQSMALTDAASRIEDADMAQEMSDFLKFKLLSEVALSMVSQANQIPQMV SELLQS FlagellinMRINTNINSMRTQEYMRQNQTKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 33RARENGLGVAANNTQDGMSLIRTADSAMNSVSNILLRMRDLANQSANGTNTD BacillusDNQKALDKEFSALKEQIDYISKNTEFNDKKLLNGENKTIAIQTLDNADTTKQ thuringiensisININLADSSTSALQIDKLTISGKTTDTTKTQTITVTDDEIKAAKTDIDEFNDAKKALADLKAESAPSKGDGSSDDEIKEAVSNFKKSFEKIQKFMNDSDIKTVQTEIEKFDAAAPALDKAKGMGIAFTSAMDPKAGTITKAATRQNASDAIKSIDAALETIASNRATLGATLNRLDFNVNNLKSQSSSMAAAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMTGITINLEIDFFAYYRFSICRKVNIKKWGFLIMRINTNINSMRTQEYMRQN SEQ ID NO: 34QTKMSNAMDRLSSGKRINNASDDAAGLAIATRMRARENGLGVAANNTQDGMS BacillusLIRTADSAMNSVSNILLRMRDLANQSANGTNTDDNQKALDKEFSALKEQIDY thuringiensisISKNTEFNDKKLLNGENKTIAIQTLDNADTTKQININLADSSTSALQIDKLT serovarISGKTTDTTKTQTITVTDDEIKAAKTDIDEFNDAKKALADLKAESAPSKGDG pondicheriensisSSDDEIKEAVSNFKKSFEKIQKFMNDSDIKTVQTEIEKFDAAAPALDKAKGMGIAFTSAMDPKAGTITKAATRQNASDAIKSIDAALETIASNRATLGATLNRLDFNVNNLKSQSSSMAAAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQ TPQMVSKLLQFlagellin B MSIMRIGTNVLSMNARQSLYENEKRMNVAMEHLATGKKLNHASDNPANIVIVSEQ ID NO: 35 TRMYARASGMRVAIRNNEDAISMLRTAEAALQTVTNILQHMRDFAIQSANGTBacillus NSNTNRDSLNKEFQSLTEPIGYIGETTEFNDLSVFDGQNRPITLDDIGHTINthuringiensis MTKHIPPSPTQHDIKISTEQEARAAIRKIEEALQNVSLHRADLGSMINRLQFserovar Berliner NIENLNSQSMALIDTASQVEDADMAQEISDFLKFKLLTAVALSVVSQANQIPQIVSKLLQS Flagellin AMARITINLEIDFFAYYRFSICRKVNIKKWGFLNMRINTNINSMRTQDYMRQN SEQ ID NO: 36QAKMSNAMDRLSSGKRINNASDDAAGLAIATRMRARESGLGVAANNTQDGMS BacillusLIRTADSAMNSVSNILLRMRDISNQSANGTNTDKNQSALDKEFAALKDQIDY thuringiensisISKNTEFNDQKLLDGSKKSIAIQTLDNADTNKQIDIQLSNVSTKELKLDTLS serovar BerlinerIEGSSSKTFTITADDMLAVGTANATAKAKAGTLKGLNVTTGDLTAAKTDVQDFRAAFDKVKGFMGSTEVTNIEKALTKFDGDQSLANAKAIGDALTSDLATTIAKDQTYSKNVSNASSAIASIDAALESIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRIGTNVLSMNARQSLYENEKRMNVAMEHLATGKKLNHASNNPANVAIVTRM SEQ ID NO: 37HARASGMRVAIRNNEDAISMLRTAEAALQTVTNVLQRMRDVAVQSANGTNSS Bacillus cereusKNRDSLNKEFQSLTEQIGYIDETTEFNDLSVFDGQNRTVTLDDIGHTVNVTK strain Q1HIPPSPTQHDINISTEQEARAAIRKIEEALQNVSLHRADLGAMINRLQFNIENLNSQSTALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQMV SKLLQS FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 38RARESGLSVAADNTQNGMSLIRTADSAMNSVSNILLRMRDIANQSANGTNTD Bacillus cereusKNQVALQKEFAALKEQITYIADNTQFNDKNLLNGNQTINIQTLDSHDSTKQI strain Q1GIDLKSATLEALGIKDLTVGAVGSTEAKNYVDAKEALAKNVAANEFIDAKKALDGNAIAKGYVEAKTAFDDAKPEVKALVSNYTDALAALAKDDTNDDLKKDVADTKALMDANTVAKTYFEAKTAHDGADQAIKDIVTTYDSKLGALDDAANKAISDFDKAKAAFDESPAAKELVKTMDDAKQAATQNNTANAYLVAKAAAELAPNDADKKAELENATKALEKDDTAKGLVKTYENAKEALNPANAMPLDAVKQIDAALKTVADNRATLGATLNRLDFNVNNLKSQSSAMAASASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRIGTNFLSMNARQSLYENEKRMNVAMEHLATGKKLNHASDNPANIAIVTRM SEQ ID NO: 39HARANGMRVAIRNNEDAISMLRTAEAALQTVMNILQRMRDLAIQSANSTNSN BacillusKNRDSLNKEFQSLTEQISYIGETTEFNDLSVFDGQNRPVTLDDIGHTVHISK thuringiensisSIPPPSPTQHDIKISTEQEARAAILKIEEALQSVSLHRADLGAMINRLHFNI serovar morrisoniENLNSQSMALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQM VSKLLQS FlagellinMRINTNINSMRTQEYMRQNQTKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 40RARENGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDIANQSANGTNTS BacillusDNQKALDKEFSALKEQIDYISKNTEFNDKKLLNGDNKSIAIQTLDNADTTKQ thuringiensisININLADSSTSALNIDKLSIEGTGNKTITLTAADIAKDKTNIDAVGTAKTAL serovarAGLTGTPAAAAINSAVADFKTAFAKADKNLMSDAQIKSVTDAITAFEADATP neoleonensisDLTKAKAIGTAYTAPAAGDITKASPNASEAIKSIDAALDTIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQ ANQTPQMVSKLLQFlagellin MTGITINLEIDFFAYYRFSICRKVNIKKWGFLNMRINTNINSMRTQEYMRQNSEQ ID NO: 41 QAKMSNAMDRLSSGKRINNASDDAAGLAIATRMRARESGLGVAANNTQDGMSBacillus LIRTADSALNSVSNILLRMRDIANQSANGTNTGDNQKALDKEFSALKEQIDYthuringiensis ISKNTEFNDKKLLNGDNKSIAIQTLDNADTAKQININLADSSTKALNIDTLSserovar morrisoni IAGTTDKTITITAKDLTDNKATLDALKTAKADLAKLDDKSDQATIDKAVDAFKTAFNNVDKNLLSDKAIEGITDKMTAFDGTHTAAAAIGTAYTEPTAGDITKSAPNASGAIKSIDAALETIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 42RARESGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDIANQSANGTNTG BacillusDNQKALDKEFSALKEQIDYISKNTEFNDKKLLNGDNKSIAIQTLDNADTAKQ thuringiensisININLADSSTKALNIDTLSIAGTTDKTITITAKDLTDNKATLDALKTAKADL serovar morrisoniAKLDDKSDQATIDKAVDAFKTAFNNVDKNLLSDKAIEGITDKMTAFDGTHTAAAAIGTAYTEPTAGDITKSAPNASGAIKSIDAALETIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQT PQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 43RARESGLGVAANNTQDGMSLIRTADSAMNSVSNILLRMRDIANQSANGTNTN BacillusGNQAALNKEFDALKQQINYISTNTEFNDKKLLDGSNKTIAIQTLDNADTSKK thuringiensisIDIQLADVSTKSLNIDKLKIGGVSKETTDAVGDTFTKLSTTATTDMGALKIE serovarVEAAMKEFDKVKGAMSAEDAKAVTDKLDAFNTAAAATNDAATIAAAKALGAA jegathesanFDKTKVEMADPNASVAAIDSALENIASNRATLGATLNRLDFNVNNLKSQQSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 44RARESGLGVAANNTQDGMALIRTADSAMNSVSNILLRMRDIANQSANGTNTD Bacillus cereusKNQAALQKEFGELQKQIDYIAGNTQFNDKNLLDGSNPSISIQTLDSADQSKQ stainISIDLKSATLEALGIKDLTVGATENTLAKATITAKDAFDAAKDASDAAKKEI ATCC 10987DAAAKDTPSKNDAQLAKEYIEAKATLATLKPTDATYAAKAAELDAATTALNDNAKVLVDGYEKKLTTTKTKEAEYTAAKEQSTKSTAAADLVTKYETAKSNALGNDIAKEYLEAKTAYEANKNDISSKSRFEAAETELNKDITANKAAKVLVETYEKAKTAGTTEKSLVAVDKIDEALKTIADNRATLGATLNRLDFNVNNLKSQSASMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMTGITINLEIDFFAYYRFSICRKVNIKKWGFLIMRINTNINSMRTQEYMRQN SEQ ID NO: 45QAKMSNAMDRLSSGKRINNASDDAAGLAIATRMRARESGLGVAANNTQDGMS BacillusLIRTADSAMNSVSNILLRMRDLANQSANGTNTNENQAALNKEFDALKEQINY thuringiensisISTNTEFNDKKLLDGSNKTIAIQTLDNADTSKKIDIKLADVSTESLKIDKLK serovarIGGVSKETTDAVSETFTKLSTTKTTDKDALKAEVEAAMKEFDKVKGAMSTED monterreyAKAVTDKLGLFNTAAAGTDDTAIATAAKNLGAAFDKTKVNMADPNASVAAIDSALENIASNRATLGATLNRLDFNVNNLKSQQSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRIGTNVLSLNARQSLYENEKRMNVAMEHLATGKKLNNASDNPANIAIVTRM SEQ ID NO: 46HARASSMRVAIRNNEDAISMLRTAEAALQTVTNVLQRMRDLAVQSANDTNSN Bacillus cereusKNRDSLNKEFQSLTEQIGYIDETTDFNDLSVFDGQNRTVTLDDIGHTVNVTK strain NC7401HIPPSPTQHDINISTEQEARAAIRKIEEALQNVSLHRADLGAMINRLQFNIENLNSQSTALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQMV SKLLQS FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 47RARESGLGVASNNTQDGMSLIRTADSALNSVSNILLRMRDLANQSANGTNTN Bacillus cereusENKAAMQKEFGELKEQIKYIAENTQFNDQHLLNADKGITKEIAIQTLDSDSD strain NC7401SKQIKIKLQGSSLEALDIKDLQIGNTELAQKDLDLLNATMDRLDATVPGTRDVDVQAAKDAFDKVKGFYTNSDSVKAIERAFEDYATASTAGTAKADAATAIKAAFDLAANKVGKPATGGAQGSANSLGAITKIDAALKTVADNRATLGATLNRLDFNVNNLKSQASSMAAAASQVEDADMAKEMSEMTKFKILNEAGISMLSQANQT PQMVSKLLQFlagellin (A-type) MRINTNINSLRTQEYMRQNQAKMSNSMDRLSSGKRINNASDDAAGLAIATRMSEQ ID NO: 48 RARESGLNVAANNTQDGMSLIRTADSALGSVSNILLRMRDLANQSANGTNTSBacillus cereus DNQAAMQKEFAELQKQITYIADNTQFNDKNLLQSNSSINIQTLDSSDGNQQIstrain AH820 GIELKSASLKSLGIEDLAIGASVNPLAKATVEASEAYDKAKADTAAFAKSIADTAATGTGAAKADAAAVDAYIKEADPTAKGNLYTGLTADQKKLADEHNTLKAAEDGKKAELTMATTKSTADGTAKGLVDAYDNAKSDAMNDPKAKAYLEAKMAYEKDTSNVANKQKLDSTKEAMEKDPASKDLVVKLDAAKAAATNGTPLDAVSKIDAALKTVADNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 49RARESGLGVASNNTQDGMSLIRTADSALNSVSNILLRMRDLANQSANGTNTN Bacillus cereusENKAAMQKEFGELKEQIKYIAENTQFNDQHLLNADKGITKEIAIQTLDSDSD AH187SKQIKIKLQGSSLEALDIKDLQIGNTELAQKDLDLLNATMDRLDATVPGTRDVDVQAAKDAFDKVKGFYTNSDSVKAIERAFEDYATASTAGTAKADAATAIKAAFDLAANKVGKPATGGAQGSANSLGAITKIDAALKTVADNRATLGATLNRLDFNVNNLKSQASSMAAAASQVEDADMAKEMSEMTKFKILNEAGISMLSQANQT PQMVSKLLQ FlagellinMDFFAYYRFSICRKVNIKKWGFFYMRINTNINSMRTQEYMRQNQAKMSNAMD SEQ ID NO: 50RLSSGKRINNASDDAAGLAIATRMRARESGLGVASNNTQDGMSLIRTADSAL Bacillus cereusNSVSNILLRMRDLANQSANGTNTNENKAAMQKEFGELKEQIKYIAENTQFNDQHLLNADKGITKEIAIQTLDSDSDSKQIKIKLQGSSLEALDIKDLQIGNTELAQKDLDLLNATMDRLDATVPGTRDVDVQAAKDAFDKVKGFYTNSDSVKAIERAFEDYATASTAGTAKADAATAIKAAFDLAANKVGKPATGGAQGSANSLGAITKIDAALKTVADNRATLGATLNRLDFNVNNLKSQASSMAAAASQVEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ Flagellin protein FlaMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 51RARESGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDIANQSANGTNTG Bacillus cereusDNQKALDKEFSALKEQIDYISKNTEFNDKKLLNGENTSIAIQTLDSADTAKQININLADSSTSALLIDKLSISGAGAGTALAGVATADINAAGTKQAALSGLTGSKTTDELDDAVKEFKTEFDKVKSGLSAENADKITAAMDKYTNNKTLDNAKAIGDLYKTMAPADSTVVGTAGTKGQALIDLNATATGDTAQKRQVAVDAFKDDFDKIKGGLNAQDAAKVTAALDKFNKADGSGNTLENAQEIGKVFAEVAAGSTKSNASDAIKSIDKALETIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQTKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 52RSREGGLNVAARNTEDGMSLIRTADSALNSVSNILLRMRDLANQSASETNTS BacillusKNQAAMQKEFDQLKEQIQYIADNTEFNDKKLLDGSNSTINIQTLDSHDKNKQ thuringiensisITISLDSASLKNLDITDLAIGSNTVNKNDLDTLNNSMKRLETAAADAAVQAQ Strain HD-771DVTDAKNAFNKVKSGYTPAEVEKMEDAFKAYDKVVADPAKTDALLKAAAEKI [51]NTEFKTLTAPTATAFDPSSSVEKIDKAIETIASSRATLGATLNRLDFNVTNLKSQENSMAASASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSK LLQ FlagellinMRINTNINSMRTQEYMRQNQTKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 53RSREGGLNVAARNTEDGMSLIRTADSALNSVSNILLRMRDLANQSASETNTS BacillusKNQAAMQKEFDQLKEQIQYIADNTEFNDKKLLDGSNSTINIQTLDSHDKNKQ thuringiensisITISLDSASLKNLDITDLAIGSNTVNKNDLDTLNNSMKRLETAAADAAVQAQ serovar sottoDVTDAKNAFNKVKSGYTPAEVEKMEDAFKAYDKVVADPAKTDALLKAAAEKI [52]NTEFKTLTAPTATAFDPSSSVEKIDKAIETIASSRATLGATLNRLDFNVTNLKSQENSMAASASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSK LLQ FlagellinMGVLNMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLA SEQ ID NO: 54IATRMRARENGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDIANQSAN BacillusGTNTGDNQKALDKEFSALKEQIDYISKNTEFNDKKLLNGDNKSIAIQTLDNA thuringiensisDTSKQINIDLANTSTSSLKIDKLSIEGKGNQTIAITAADIAKDTNIAALTSA serovarQGKLAALTGTPAPAALTTAVDEFKAAFEKVDKNLMSDTQITGIENAIKAYDG NovosibirskATTKTLALAQAVGTAYTAPTPGDITKELPNASSSIKSIDAALETIASNRATLGATLNRLDFNVNNLKSQASSMASAASQIEDADMAKEMSEMTKFKILNEAGIS MLSQANQTPQMVSKLLQFlagellin MGVLNMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLASEQ ID NO: 55 IATRMRARESGLGVAANNTQDGISLIRTADSAMNSVSNILLRMRDLANQSANBacillus GTNTSENQAALDKEFGALKEQINYISTNTEFNDKKLLDGSNETIAIQTLDNAthuringiensis DEGKKIDIKLANVSTDSLKIDKLTIGGAAQKTVDAVADKFNALKTTTTTDKAserovar londrina AIQTEVDAVMKEFDKVKGSMSAEDAKVITDKLKDYNDAADTDTAKATAAKDLGAAFDKTKVNIANPNAAVAAIDSALENIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRIGTNVLSMNARQSLYENEKRMNVAMEHLATGKKLNHASNNPANIAIVTRM SEQ ID NO: 56HARASGMRVAIRNNEDALSMLRTAEAALQTVTNILQRMRDLAVQSANVTNSN Bacillus cereusKNRNSLNKEFQSLTEQISYIGETTEFNDLSVFDGQNRPVTLDDIGYTVNVTK strain E33LHTPPSPTQHDIKISTEQEARAAIRKIEEALQNVSLHRADLGSMMNRLQFNIENLNSQSMALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQMV SKLLQS FlagellinMRINTNINSMRTQEYMRQNQAKMSTAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 57RARESGLGVAANNTQDGISLIRTADSAMNSVSNILLRMRDLANQSANGTNTD Bacillus cereusKNQGALDKEFAALKEQIDYISKNTEFNDKKLLDGSNKAIAIQTLDSDDKGKQ strain E33LIDISLSDTSTTALKINNLSIAANGLGIGSGKELVGVADNTIANASAEALKKLDGTTGDTDVKRSNAVKAFTDQYKDLKVAMNAKDVETIDAAIKKFEGANTLENAQAIGAAFEGAAKATLTTDINNATLTSKALSDLDTDSTTETRKAAMKDFVAAFDKVKGSMNSSDVTKISDAIDRFSKTDDSGNTLEAARAIGDAFKAATTNGKTSTATDANSAIKAIDEALETIASNRATLGATLNRLDFNVNNLKNQASSMASAASQVEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSTAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 58RARESGLGVAANNTQDGISLIRTADSAMNSVSNILLRMRDLANQSANGTNTD Bacillus cereusKNQAALDKEFNALKEQIDYISKNTEFNDKKLLDGSNKSIAVQTLDNADTSKQ strain FRI-35ININLSNTSTKALEINSLTISGTTPIAGKNETSKITAEQMTAASDALEKFKTAQEGLANLTEPTKGSDGKPEAGTGSSNEDIVKAVKAFKEAFKNIQPLMSDTDITTVQNKIDLFDEDAPDLSAAKLIGTTFEESMKPVADKEITKAAVKPNASDAIAAIDAALTKVADNRATLGATLNRLDFNVNNLKSQASSMASAASQVEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRIGTNVLSLNARQSLYENEKRMNVAMEHLATGKKLNNASDNPANIAIVTRM SEQ ID NO: 59HARASGMRVAIRNNEDAISMLRTAEAALQTVTNVLQRMRDLAVQSANGTNSN Bacillus cereusKNRDSLNKEFQSLTEQIGYIDETTEFNNLSVFDGQNRPVTLDDIGHTVNVTK strain FRI-35HIPPFPTQHDINISTEQEARAAIRKIEEALQNVSLHRADLGAMINRLQFNIENLNSQSTALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQVPQMV SKLLQS FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 60RAHESGLSVAARNTSDGISLIRTADSALQSVSNILLRMRDIANQTANGTNKD BacillusTDIEALGKEFAALKEQITYVSDNTKFNGRELLKGGDDINIQTYDGSDESQQI thuringiensisKIKISELDLSSLDTGEVTDSDTARGTVSTLDDAITNIASKRAELGATLNRLDYNTQNVNSEAASMAASASQIEDADMAKEMSEMTKFKILSEAGISMLSQANQT PQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 61RAHESGLSVAARNTSDGISLIRTADSALQSVSNILLRMRDIANQTANGTNKD Bacillus cereusTDIEALGKEFAALKEQITYVSDNTKFNGRELLKGGDDINIQTYDGSDESQQI strain ATCC 4342KIKISELDLSSLDTGEVTDSDTARGTVSTLDDAITNIASKRAELGATLNRLDYNTQNVNSEAASMAASASQIEDADMAKEMSEMTKFKILSEAGISMLSQANQT PQMVSKLLQ FlagellinMRIGTNFLSMNARQSLYENEKRMNVAMEHLATGKKLNHASDNPANIAIVTRM SEQ ID NO: 62HARANGMRVAIRNNEDAISMLRTAEAALQTVMNILQRMRDLAIQSANSTNSN BacillusKNRDSLNKEFQSLTEQISYIGETTEFNDLSVFDGQNRPVTLDDIGHTVHISK thuringiensisSIPPPSPTQHDIKISTEQEARAAILKIEEALQSVSLHRADLGAMINRLHFNIENLNSQSMALTDAASRIEDADMAQEMSDFLKFKLLTEVALSMVSQANQIPQM VSKLLQS FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 63RARESGLGVAANNTQDGMSLIRTADSALNSVSNILLRMRDIANQSANGTNTG BacillusDNQKALDKEFSALKEQIDYISKNTEFNDKKLLNGDNKSIAIQTLDNADTAKQ thuringiensisININLADSSTKALNIDTLSIAGTTDKTITITAKDLTDNKATLDALKTAKADLAKLDDKSDQATIDKAVDAFKTAFNNVDKNLLSDKAIEGITDKMTAFDGTHTAAAAIGTAYTEPTAGDITKSAPNASGAIKSIDAALETIASNRATLGATLNRLDFNVNNLKSQSSSMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQT PQMVSKLLQ FlagellinMRINHNITALNTYRQFNNANNAQAKSMEKLSSGQRINSASDDAAGLAISEKM SEQ ID NO: 64RGQIRGLDQASRNAQDGVSLIQTAEGALNETHDILQRMRELVVQAGNGTNKT Bacillus EDLDAIQDEIGSLIEEIGGETDSKGISDRAQFNGRNLLDGSLDITLQVGANA aryabhattaiGQQVNLKIGDMSAGALGADTDSDGAADAFVNSINVKDFATTSFDDQLAIIDGAINQVSEQRSGLGATQNRLDHTINNLSTSSENLTASESRIRDVDYALAA FlagellinMRINTNINSMRTQEYMRQNQDKMNTSMNRLSSGKQINSASDDAAGLAIATRM SEQ ID NO: 65RAKEGGLNVGAKNTQDGMSALRTMDSALNSVSNILLRMRDLATQSATGTNQG BacillusNDRESLDLEFQQLTEEITHIAEKTNFNGNALLSGSGSAINVQLSDAAEDKLT manliponensisIAAIDATASTLLKGAVDVKTEDKADAAITKIDQAIQDIADNRATYGSQLNRLDHNLNNVNSQATNMAAAASQIEDADMAKEMSEMTKFKILSEAGVSMLSQANQ TPQMVSKLLQFlagellin MRIGSWTATGMSIVNHMNRNWNAASKSMLRLSSGYRINSAADDAAGLAISEKSEQ ID NO: 66 MRGQIRGLTMASKNIMDGVSLIQTAEGALNETHAIVQRMRELAVQAATDTNTLysinibacillus DDDRAKLDLEFQELKKEIDRISTDTEFNTRTLLNGDYKDNGLKIQVGANSGQsp. strain BF-4 AIEVKIGDAGLAGIGLSTESIATREGANAALGKLDEATKNVSMERSRLGAYQNRLEHAYNVAENTAINLQDAESRIRDVDIAKEMMNMVKSQILAQVGQQVLAM HMQQAQGILRLLGFlagellin MKIGSWTATGMSIVNHMNRNWNAASKSMLRLSSGYRINSAADDAAGLAISEKSEQ ID NO: 67 MRGQIRGLTMASKNIMDGVSLIQTAEGALNETHAIVQRMRELAVQAATDTNTLysinibacillus DDDRAKLDLEFQELKKEIDRISTDTAFNTRTLLNGDYKDNGLKIQVGANSGQsp. strain AIEVKIGDAGLAGIGLSTESIATREGANAALGKLDEATKNVSMERSRLGAYQ13S34_air NRLEHAYNVAENTAINLQDAESRIRDVDIAKEMMHMVKSQILAQVGQQVLAMHIQQAQGILRLLG FlagellinMIISHNLTALNTMNKLKQKDLAVSKSLGKLSSGLRINGASDDAAGLAISEKM SEQ ID NO: 68RGQIRGLNQASRNIQDGISLIQVADGAMQEIHSMLQRMNELAVQASNGTYSG PaenibacillusSDRLNIQSEVEQLIEEIDEIAGNTGFNGIKLLNGNNEKTEKTEKTGSVVSVN sp. strainNPPNNKLITISSPVGTSVSEILNNLLTVFNEAKNGQVGDSDSKRVSSKFTLS HW567INNDELSIVCDTGDGFLLSGGSPNLFYQGYIGGSYKYKFTEFINENDFINIMDIGGANGGDTLKFNFSSISKEPEEQKEQKGLTLQIGANSGETLNIKLPNVTTSAIGISSIDVSTIPNAESSLSSISAAIDKVSAERARMGAYQNRLEHSRNNVVTYAENLTAAESRIRDVDMAKEMMELMKNQIFTQAGQAMLLQTNTQPQAILQL LK FlagellinMRINTNINSMRTQEYMRQNQAKMSNAMDRLSSGKRINNASDDAAGLAIATRM SEQ ID NO: 69RARESGLGVAANNTQDGMSLIRTADSAMNSVSNILLRMRDLANQSANGTNTK BacillusENQDALDKEFGALKEQIDYISKNTEFNDKKLLNGDNKSIAIQTLDNADTAKQ anthracisININLADSSTKALNIDSLTISGSKDATITITAEDITAASAEITAAKGARTALANLKDTPADPTKDPAASTPAEIKAAVDDFKGKFEKIKGLMNDTDVKAVEEKIKEFETTSTLAKAQAIGTAFTTGMEPKAGNITKNVPAASSSIKAIDSALETIASNRATLGATLNRLDFNVNNLKSQSSAMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMQKSQYKKMGVLKMRINTNINSMRTQEYMRQNQDKMNVSMNRLSSGKRINSA SEQ ID NO: 70ADDAAGLAIATRMRARQSGLEKASQNTQDGMSLIRTAESAMNSVSNILTRMR BacillusDIAVQSSNGTNTAENQSALQKEFAELQEQIDYIAKNTEFNDKNLLAGTGAVT anthracisIGSTSISGAEISIETLDSSATNQQITIKLANTTAEKLGIDATTSNISISGAASALAAISALNTALNTVAGNRATLGATLNRLDRNVENLNNQATNMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQDKMNVSMNRLSSGKRINSAADDAAGLAIATRM SEQ ID NO: 71RARQSGLEKASQNTQDGMSLIRTAESAMNSVSNILTRMRDIAVQSSNGTNTA BacillusENQSALQKEFAELQEQIDYIAKNTEFNDKNLLAGTGAVTIGSTSISGAEISI anthracisETLDSSATNQQITIKLANTTAEKLGIDATTSNISISGAASALAAISALNTALNTVAGNRATLGATLNRLDRNVENLNNQATNMASAASQIKDADKAKEMSEMTKFKILNEAGISMLSQANQTPQMVSKLLQ FlagellinMRINTNINSMRTQEYMRQNQDKMNVSMNRLSSGKRINSAADDAAGLAIATRM SEQ ID NO: 72RARQSGLEKASQNTQDGMSLIRTAESAMNSVSNILTRMRDIAVQSSNGTNTA BacillusENQSALQKEFAELQEQIDYIAKNTEFNDKNLLAGTGAVTIGSTSISGAEISI anthracisETLDSSATNQQITIKLANTTAEKLGIDATTSNISISGAASALAAISALNTALNTVAGNRATLGATLNRLDRNVENLNNQATNMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMV FlagellinMNVSMNRLSSGKRINSAADDAAGLAIATRMRARQSGLEKASQNTQDGMSLIR SEQ ID NO: 73TAESAMNSVSNILTRMRDIAVQSSNGTNTAENQSALQKEFAELQEQIDYIAK BacillusNTEFNDKNLLAGTGAVTIGSTSISGAEISIETLDSSATNQQITIKLANTTAE anthracisKLGIDATTSNISISGAASALAAISALNTALNTVAGNRATLGATLNRLDRNVE strain H9401NLNNQATNMASAASQIEDADMAKEMSEMTKFKILNEAGISMLSQANQTPQMV SKLLQ FlagellinMRINHNITALNTYRQFNNANNAQAKSMEKLSSGQRINSASDDAAGLAISEKM SEQ ID NO: 74RGQIRGLDQASRNAQDGVSLIQTAEGALNETHDILQRMRELVVQAGNGTNKT BacillusEDLDAIQDEIGSLIEEIGGEADSKGISDRAQFNGRNLLDGSLDITLQVGANA megateriumGQQVNLKIGDMSAGALGADTNSDGAADAFVNSINVKDFTATSFDDQLAIIDG strain WSH-002AINQVSEQRSGLGATQNRLDHTINNLSTSSENLTASESRIRDVDYALAA FlagellinMRINHNLPALNAYRNLAQNQIGTSKILERLSSGYRINRASDDAAGLAISEKM SEQ ID NO: 75RGQIRGLEQGQRNTMDGVSLIQTAEGALQEIHEMLQRMRELAVQAANGTYSD AneurinibacillusKDKKAIEDEINQLTAQIDQIAKTTEFNGIQLIGDSDSTSLQDVKIQYGPKKE sp. XH2DSLTLELTTQPEADPPFAAGCKADKASLKIDNVDVISDPEGAIETFKAAIDQVSRIRSYFGAIQNRLEHVVNNLSNYTENLTGAESRIRDADMAKEMTEFTRFNIINQSATAMLAQANQLPQGVLQLLKG

N- and C-Terminal Conserved Regions of Flagellin

The flagellin or flagellin-associated polypeptide can comprise atruncated N-terminal polypeptide and an amino acid sequence of thetruncated N-terminal polypeptide can comprise SEQ ID NO: 76, 78, 80, 82,84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 109, 110, 112,114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138, 140,142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166, 168,170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194, 196,198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224,752, or any combination thereof.

The flagellin or flagellin-associated polypeptide can comprise atruncated C-terminal polypeptide and an amino acid sequence of thetruncated C-terminal polypeptide can comprise SEQ ID NO: 77, 79, 81, 83,85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113, 115,117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 141, 143, 145,147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173,175, 177, 179, 181, 183, 185, 187, 189, 191, 201, 203, 205, 207, 209,211, 213, 215, 217, 219, 221, 223, 225, or any combination thereof.

N-terminal and C-terminal conserved regions were identified from fulllength flagellin sequences from diverse strains of Bacillus spp. andother Eubacteria (Table 2). Conserved N- and C-terminal domains wereidentified using BLAST multiple alignment software and assignedfunctional annotations based on individual hits searching againstBacillus and other Eubacterial bacterial databases. The start site forthe N-terminal region of the coding sequences is bolded methionine (M).The conserved domains are provided as amino acid sequences N-terminus(left column) and C-terminus (right column).

TABLE 2 N- and C-terminal conserved regions of flagellins SEQ ID NO:Conserved N-terminus Conserved C-terminus Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM IDAAITTVAGQRATLGATLNRFE N-SEQ ID NO: 76SNAMDRLSSGKRINSASDDAAGLAIATRM FNANNLKSQETSMADAASQIE C-SEQ ID NO: 77KAREGGLNVAGRNTQDGMSLIRTADSALN DADMAKEMSEMTKFKILNEAGBacillus thuringensis SVSNILLRMRDLANQSANGTNTKGNQASL ISMLSQANQTPQMVSKLLQstrain 4Q7 QKEFAQLTEQIDYIAKNTQFNDQQLLGTAD [CDS of SEQ ID NO: 1] KKIKIQTLFlagellin GFLN

RINTNINSMRTQEYMRQNQAKM IDAAITTVAGQRATLGATLNRFE N-SEQ ID NO: 78SNAMDRLSSGKRINSASDDAAGLAIATRM FNANNLKSQETSMADAASQIE C-SEQ ID NO: 79KAREGGLNVAGRNTQDGMSLIRTADSALN DADMAKEMSEMTKFKILNEAGBacillus thuringiensis, SVSNILLRMRDLANQSANGTNTKGNQASLISMLSQANQTPQMVSKLLQ strain HD1002 QKEFAQLTEQIDYIAKNTQFNDQQLLGTAD[CDS of SEQ ID NO: 2] KKIKIQTL Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM IDAAITTVAGQRATLGATLNRFE N-SEQ ID NO: 80SNAMDRLSSGKRINSASDDAAGLAIATRM FNANNLKSQETSMADAASQIE C-SEQ ID NO: 81KAREGGLNVAGRNTQDGMSLIRTADSALN DADMAKEMSEMTKFKILNEAGBacillus thuringiensis, SVSNILLRMRDLANQSANGTNTKGNQASLISMLSQANQTPQMVSKLLQ strain HD-789 QKEFAQLTEQIDYIAKNTQFNDQQLLGTAD[CDS of SEQ ID NO: 3] KKIKIQTL Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM IDAAITTVAGQRATLGATLNRFE N-SEQ ID NO: 82SNAMDRLSSGKRINSASDDAAGLAIATRM FNANNLKSQETSMADAASQIE C-SEQ ID NO: 83KAREGGLNVAGRNTQDGMSLIRTADSALN DADMAKEMSEMTKFKILNEAG Bacillus cereusSVSNILLRMRDLANQSANGTNTKGNQASL ISMLSQANQTPQMVSKLLQ strain G9842QKEFAQLTEQIDYIAKNTQFNDQQLLGTAD [CDS of SEQ ID NO: 4] KKIKIQTL FlagellinGFLN

RINTNINSMRTQEYMRQNQAKM QLDAALTKVADNRATLGATLNR N-SEQ ID NO: 84SNSMDRLSSGKRINSAADDAAGLAIATRM LDFNVNNLKSQENSMAASASQ C-SEQ ID NO: 85KAREGGLNVAARNTQDGMSLIRTADSALN IEDADMAKEMSEMTKFKILNEABacillus thuringiensis SVSNILLRMRDLANQSATGTNTTKNQVALGISMLSQANQTPQMVSKLLQ serovar indiana  NKEFAALKEQITYIADNTQFNDKNLLKSTQstrain HD521 EIKIQTL [CDS of SEQ ID NO: 5] Flagellin WGFLI

RINTNINSMRTQEYMRQNQAK AIAAIDAALTKVADNRATLGATL N-SEQ ID NO: 86MSNSMDRLSSGKRINNASDDAAGLAIATR NRLDFNVNNLKSQSSSMASAA C-SEQ ID NO: 87MRARESGLGVAADNTQNGMSLIRTADSA SQIEDADMAKEMSEMTKFKILNBacillus thuringiensis MNSVSNILLRMRDIANQSANGTNTNENKSEAGISMLSQANQTPQMVSKLL strain CTC ALQKEFAQLQKQITYIAENTQFNDKNLLNE Q[CDS of SEQ ID NO: 6] DSEVKIQTLDS Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVNNLKSQS N-SEQ ID NO: 88SNAMDRLSSGKRINNASDDAAGLAIATRM SSMAAAASQIEDADMAKEMSE C-SEQ ID NO: 89RARENGLGVAANNTQDGMSLIRTADSAM MTKFKILNEAGISMLSQAN BacillusNSVSNILLRMRDLANQSANGTNTDDNQK thuringiensis ALDKEFSALKEQIDYISKNTEFNDKKLLserovar yunnanensis strain IEBC-T20001 [CDS of SEQ ID NO: 7] FlagellinGFLN

RINTNINSMRTQEYMRQNQAKM IDAALKTVADNRATLGATLNRL N-SEQ ID NO: 90SNAMDRLSSGKRINNASDDAAGLAIATRM DFNVNNLKSQSASMASAASQIE C-SEQ ID NO: 91RARENGLGVAANNTQDGMSLIRTADSAL DADMAKEMSEMTKFKILNEAGBacillus thuringiensis QSVSNILLRMRDLANQSANGTNTDENKAA ISMLSQANQTPQMVSKLLQserovar tolworthi MEKEFGQLKDQIKYITDNTQFNDKNLLDA [CDS of SEQ ID NO: 8]Flagellin MGVLN

RINTNINSMRTQEYMRQNQAK RATLGATLNRLDFNVNNLKSQQ N-SEQ ID NO: 92MSNSMDRLSSGKRINNASDDAAGLAIATR SSMASAASQVEDADMAKEMS C-SEQ ID NO: 93MRARESGLGVAANNTQDGMSLIRTADSA EMTKFKILNEAGISMLSQANQT Bacillus cereusMNSVSNILLRMRDIANQSANGTNTDKNQ PQMVSKLLQ strain FM1VALQKEFGELQKQIDYIAKNTQFND [CDS of SEQ ID NO: 9] Flagellin MGVLN

RIGTNVLSMNARQSFYENEKR RADLGAMINQLQFNIENLNSQS N-SEQ ID NO: 94MNVAIEHLATGKKLNHASDNPANVAIVTR TALTDAASRIEDADMAQEMSD C-SEQ ID NO: 95MHARTSGIHVAIRNNEDAISMLRTAEAAL FLKFKLLTEVALSMVSQANQIP Bacillus cereusQTVTNILQRMRDVAVQSANGTNSNKNRD QMVYKLLQ strain FM1SLNKEFQSLTEQIGYIDETTEFND [CDS of SEQ ID NO: 10] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM AVDSIDAALKTVASNRATLGATL N-SEQ ID NO: 96SNAMDRLSSGKRINNASDDAAGLAIATRM NRLDFNVNNLKSQSASMASAA C-SEQ ID NO: 97RARESGLGVAANNTQDGMSLIRTADSALN SQIEDADMAKEMSEMTKFKILNBacillus thuringiensis SVSNILLRMRDIANQSANGTNTADNQQALEAGISMLSQANQTPQMVSKLL strain MC28 QKEFGQLKEQISYIADNTEFNDKTLL Q[CDS of SEQ ID NO: 11] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM LGATLNRLDFNVTNLKSQENS N-SEQ ID NO: 98SNSMDRLSSGKRINNASDDAAGLAIATRM MAASASQIEDADMAKEMSEM C-SEQ ID NO: 99RSREGGLNVAARNTEDGMSLIRTADSALN TKFKILNEAGISMLSQANQTPQBacillus bombysepticus SVSNILLRMRDLANQSASGTNTDKNQAA MVSKLLQ strain WangMQKEFDQLKEQIQYI [CDS of SEQ ID NO: 12] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVTNLKSQE N-SEQ ID NO: 100SNSMDRLSSGKRINNASDDAAGLAIATRM NSMAASASQIEDADMAKEMSE C-SEQ ID NO: 101RSREGGLNVAARNTEDGMSLIRTADSALN MTKFKILNEAGISMLSQANQTPBacillus thuringiensis SVSNILLRMRDLANQSASGTNTDKNQAA QMVSKLLQserovar kenyae MQKEFDQLKEQIQYI [CDS of SEQ ID NO: 13] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVTNLKSQE N-SEQ ID NO: 102SNSMDRLSSGKRINNASDDAAGLAIATRM NSMAASASQIEDADMAKEMSE C-SEQ ID NO: 103RSREGGLNVAARNTEDGMSLIRTADSALN MTKFKILNEAGISMLSQANQTPBacillus thuringiensis SVSNILLRMRDLANQSASGTNTDKNQAA QMVSKLLQserovar kenyae MQKEFDQLKEQIQYI [CDS of SEQ ID NO: 14] Flagellin (A-type)GFLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVNNLKSQS N-SEQ ID NO: 104SNAMDRLSSGKRINNASDDAAGLAIATRM SSMASAASQIEDADMAKEMSE C-SEQ ID NO: 105RARENGLGVAANNTQDGMSLIRTADSAL MTKFKILNEAGISMLSQANQTP Bacillus cereusNSVSNILLRMRDLANQSANGTNTGDNQK QMVSKLLQ [CDS of SEQ ID NO: 15]ALDKEFSALKEQIDYISKNTEFNDKKLL Flagellin (A-type) GFLN

RIGTNVLSMNARQSLYENEKRMN RADLGSMINRLQFNIENLNSQS N-SEQ ID NO: 106VAMEHLATGKKLNNASDNPANIAIVTRMH MALTDAASRIEDADMAQEMS C-SEQ ID NO: 107ARASGMRLAIRNNEDTISMLRTAEAALQTL DFLKFKLLTEVALSMVSQANQIP Bacillus cereusTNILQRMRDLAVQSANGTNSNKNRDSLNK QMVSKLLQ [CDS of SEQ ID NO: 16]EFQSLTEQIGYIGETTEFND Flagellin GVLN

RINTNINSMRTQEYMRQNQAKM AIDAALTKVADNRATLGATLNR N-SEQ ID NO: 108SNAMDRLSSGKRINNASDDAAGLAIATRM LDFNVNNLKSQSSSMASAASQI C-SEQ ID NO: 109RARESGLNVAADNTQNGMSLIRTADSAM EDADMAKEMSEMTKFKILNEABacillus thuringiensis NSVSNILLRMRDIANQSANGTNTDSNKSAGISMLSQANQTPQMVSKLLQ serovar finitimus LQKEFAELQKQITYIADNTQFNDKNLLKEDSstrain YBT-020 EVKIQTLDS [CDS of SEQ ID NO: 17] Flagellin GVLN

RINTNINSMRTQEYMRQNQAKM AAIDAALTKVADNRATLGATLN N-SEQ ID NO: 110SNAMDRLSSGKRINNASDDAAGLAIATRM RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 111RARESGLNVAADNTQNGMSLIRTADSAM QIEDADMAKEMSEMTKFKILNBacillus thuringiensis NSVSNILLRMRDIANQSANGTNTDSNKSAEAGISMLSQANQTPQMVSKLL serovar finitimus LQKEFAELQKQITYIADNTQFNDKNLLKEDSQ strain YBT-020 EVKIQTLDS [CDS of SEQ ID NO: 18] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM TVADNRATLGATLNRLDFNVN N-SEQ ID NO: 112SNAMDRLSSGKRINNASDDAAGLAIATRM NLKSQSASMASAASQIEDADM C-SEQ ID NO: 113RARESGLGVAANNTQDGMSLIRTADSALN AKEMSEMTKFKILNEAGISMLS Bacillus cereusSVSNILLRMRDLANQSANGTNTAENKAA QANQTPQMVSKLLQ stain B4264MQKEFGELKDQIKYISENTQFNDQHLL [CDS of SEQ ID NO: 19] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM AIKSIDAALDTIASNRATLGATLN N-SEQ ID NO: 114SNAMDRLSSGKRINNASDDAAGLAIATRM RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 115RARESGLGVAANNTQDGMSLIRTADSALN QIEDADMAKEMSEMTKFKILNBacillus thuringiensis SVSNILLRMRDIANQSANGTNTSDNQKALEAGISMLSQANQTPQMVSKLL serovar nigeriensis DKEFSALKEQIDYISKNTEFNDKKLL Q[CDS of SEQ ID NO: 20] Flagellin WGFLI

RINTNINSMRTQEYMRQNQAK AVDAIDAALKTVASNRATLGAT N-SEQ ID NO: 116MSNAMDRLSSGKRINNASDDAAGLAIATR LNRLDFNVNNLKSQSASMASA C-SEQ ID NO: 117MRARESGLGVAANNTQDGMSLIRTADSA ASQIEDADMAKEMSEMTKFKILBacillus thuringiensis LNSVSNILLRMRDIANQSANGTNTADNQQNEAGISMLSQANQTPQMVSKL [CDS of SEQ ID NO: 21] ALQKEFGQLKEQISYIADNTEFND LQFlagellin WGFLI

RINTNINSMRTQEYMRQNQAK AVDAIDAALKTVASNRATLGAT N-SEQ ID NO: 118MSNAMDRLSSGKRINNASDDAAGLAIATR LNRLDFNVNNLKSQSASMASA C-SEQ ID NO: 119MRARESGLGVAANNTQDGMSLIRTADSA ASQIEDADMAKEMSEMTKFKILBacillus thuringiensis LNSVSNILLRMetRDIANQSANGTNTADNNEAGISMLSQANQTPQMVSKL serovar konkukian QQALQKEFGQLKEQISYIADNTEFNDKTLLLQ strain 97-27 [CDS of SEQ ID NO: 22] Flagellin WGFLI

RINTNINSMRTQEYMRQNQAK AIASIDAALESIASNRATLGATLN N-SEQ ID NO: 120MSNAMDRLSSGKRINNASDDAAGLAIATR RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 121MRARESGLGVAANNTQDGMSLIRTADSA QIEDADMAKEMSEMTKFKILNBacillus thuringiensis MNSVSNILLRMRDISNQSANGTNTDKNQSEAGISMLSQANQTPQMVSKLL serovar konkukian ALDKEFAALKDQIDYISKNTEFNDQKLL Qstrain 97-27 [CDS of SEQ ID NO: 23] Flagellin protein FlaA GFLN

RINTNINSMRTQEYMRQNQAKM AIASIDAALESIASNRATLGATLN N-SEQ ID NO: 122SNAMDRLSSGKRINNASDDAAGLAIATRM RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 123RARESGLGVAANNTQDGMSLIRTADSAM QIEDADMAKEMSEMTKFKILNBacillus thuringiensis NSVSNILLRMRDISNQSANGTNTDKNQSAEAGISMLSQANQTPQMVSKLL serovar thuringiensis LDKEFAALKDQIDYISKNTEFNDQKLLQ strain IS5056 [CDS of SEQ ID NO: 24] Flagellin protein FlaA GFLN

RINTNINSMRTQEYMRQNQAKM AIASIDAALESIASNRATLGATLN N-SEQ ID NO: 124SNAMDRLSSGKRINNASDDAAGLAIATRM RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 125RARESGLGVAANNTQDGMSLIRTADSAM QIEDADMAKEMSEMTKFKILNBacillus thuringiensis NSVSNILLRMRDISNQSANGTNTDKNQSAEAGISMLSQANQTPQMVSKLL serovar thuringiensis LDKEFAALKDQIDYISKNTEFNDQKLLQ strain IS5056 [CDS of SEQ ID NO: 25] Flagellin B GFLN

RINTNINSMRTQEYMRQNQAKM AIASIDAALESIASNRATLGATLN N-SEQ ID NO: 126SNAMDRLSSGKRINNASDDAAGLAIATRM RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 127RARESGLGVAANNTQDGMSLIRTADSAM QIEDADMAKEMSEMTKFKILNBacillus thuringiensis NSVSNILLRMRDISNQSANGTNTDKNQSAEAGISMLSQANQTPQMVSKLL strain Bt407 LDKEFAALKDQIDYISKNTEFNDQKLL Q[CDS of SEQ ID NO: 26] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM AIASIDAALESIASNRATLGATLN N-SEQ ID NO: 128SNAMDRLSSGKRINNASDDAAGLAIATRM RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 129RARESGLGVAANNTQDGMSLIRTADSAM QIEDADMAKEMSEMTKFKILNBacillus thuringiensis NSVSNILLRMRDISNQSANGTNTDKNQSAEAGISMLSQANQTPQMVSKLL serovar chinensis  LDKEFAALKDQIDYISKNTEFNDQKLL QCT-43 [CDS of SEQ ID NO: 27] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVNNLKSQS N-SEQ ID NO: 130SNAMDRLSSGKRINNASDDAAGLAIATRM SSMASAASQIEDADMAKEMSE C-SEQ ID NO: 131RARESGLGVAANNTQDGISLIRTADSAMN MTKFKILNEAGISMLSQANQTPBacillus thuringiensis SVSNILLRMRDLANQSANGTNTNENQAAL QMVSKLLQserovar Canadensis NKEFDALKEQIDYISTNTEFNDKKLL [CDS of SEQ ID NO: 28]Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVNNLKSQS N-SEQ ID NO: 132SNAMDRLSSGKRINNASDDAAGLAIATRM SSMASAASQIEDADMAKEMSE C-SEQ ID NO: 133RARESGLGVAANNTQDGISLIRTADSAMN MTKFKILNEAGISMLSQANQTPBacillus thuringiensis SVSNILLRMRDLANQSANGTNTNENQAAL QMVSKLLQserovar galleriae NKEFDALKEQIDYISTNTEFNDKKLL [CDS of SEQ ID NO: 29]Flagellin N-terminal GVLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVNNLKSQS helical regionSNAMDRLSSGKRINNASDDAAGLAIATRM SSMASAASQIEDADMAKEMSE N-SEQ ID NO: 134RARESGLSVAANNTQDGMSLIRTADSAM MTKFKILNEAGISMLSQANQTP C-SEQ ID NO: 135NSVSNILLRMRDLSNQSANGTNTDENQQ QMVSKLLQ BacillusALNKEFAALKDQIDYISKNTEFNDKKLL weihenstephanensis [CDS of SEQ ID NO: 30]Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM IDAALETIASNRATLGATLNRLD N-SEQ ID NO: 136SNAMDRLSSGKRINNASDDAAGLAIATRM FNVNNLKSQSSSMASAASQIED C-SEQ ID NO: 137RARESGLGVAANNTQDGMSLIRTADSALN ADMAKEMSEMTKFKILNEAGISBacillus thuringiensis SVSNILLRMRDIANQSANGTNTGDNQKAL MLSQANQTPQMVSKLLQSserovar ostriniae DKEFSALKEQIDYISKNTEFNDKKLL [CDS of SEQ ID NO: 31]Flagellin WGFLI

RINTNINSMRTQEYMRQNQTK LGATLNRLDFNVNNLKSQSSSM N-SEQ ID NO: 138MSNAMDRLSSGKRINNASDDAAGLAIATR AAAASQIEDADMAKEMSEMT C-SEQ ID NO: 139MRARENGLGVAANNTQDGMSLIRTADSA KFKILNEAGISMLSQANQTPQMBacillus thuringiensis MNSVSNILLRMRDLANQSANGTNTDDNQ VSKLLQ[CDS of SEQ ID NO: 32] KALDKEFSALKEQIDYISKNTEFNDKKLL Flagellin WGFLI

RINTNINSMRTQEYMRQNQTK LGATLNRLDFNVNNLKSQSSSM N-SEQ ID NO: 140MSNAMDRLSSGKRINNASDDAAGLAIATR AAAASQIEDADMAKEMSEMT C-SEQ ID NO: 141MRARENGLGVAANNTQDGMSLIRTADSA KFKILNEAGISMLSQANQTPQMBacillus thuringiensis MNSVSNILLRMRDLANQSANGTNTDDNQ VSKLLQ[CDS of SEQ ID NO: 33] KALDKEFSALKEQIDYISKNTEFNDKKLL Flagellin WGFLI

RINTNINSMRTQEYMRQNQTK RATLGATLNRLDFNVNNLKSQS N-SEQ ID NO: 142MSNAMDRLSSGKRINNASDDAAGLAIATR SSMAAAASQIEDADMAKEMSE C-SEQ ID NO: 143MRARENGLGVAANNTQDGMSLIRTADSA MTKFKILNEAGISMLSQANQTPBacillus thuringiensis MNSVSNILLRMRDLANQSANGTNTDDNQ QMVSKLLQserovar pondicheriensis KALDKEFSALKEQIDYISKNTEFNDKKLL[CDS of SEQ ID NO: 34] Flagellin B GFLN

RINTNINSMRTQDYMRQNQAKM AIASIDAALESIASNRATLGATLN N-SEQ ID NO: 144SNAMDRLSSGKRINNASDDAAGLAIATRM RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 145RARESGLGVAANNTQDGMSLIRTADSAM QIEDADMAKEMSEMTKFKILNBacillus thuringiensis NSVSNILLRMRDISNQSANGTNTDKNQSAEAGISMLSQANQTPQMVSKLL serovar Berliner LDKEFAALKDQIDYISKNTEFNDQKLL Q[CDS of SEQ ID NO: 35] Flagellin A GFLN

ARITINLEIDFFAYYRFSICRKVNIKK AIASIDAALESIASNRATLGATLN N-SEQ ID NO: 146WGFLNMRINTNINSMRTQDYMRQNQAK RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 147MSNAMDRLSSGKRINNASDDAAGLAIATR QIEDADMAKEMSEMTKFKILNBacillus thuringiensis MRARESGLGVAANNTQDGMSLIRTADSAEAGISMLSQANQTPQMVSKLL serovar Berliner MNSVSNILLRMRDISNQSANGTNTDKNQS Q[CDS of SEQ ID NO: 36] ALDKEFAALKDQIDYISKNTEFNDQKLL Flagellin GVLY

RINTNINSMRTQEYMRQNQAKM TVADNRATLGATLNRLDFNVN N-SEQ ID NO: 148SNAMDRLSSGKRINNASDDAAGLAIATRM NLKSQSSAMAASASQIEDADM C-SEQ ID NO: 149RARESGLSVAADNTQNGMSLIRTADSAM AKEMSEMTKFKILNEAGISMLS Bacillus cereusNSVSNILLRMRDIANQSANGTNTDKNQVA QANQTPQMVSKLLQ strain Q1LQKEFAALKEQITYIADNTQFNDKNLLNGN [CDS of SEQ ID NO: 37] QTINIQTLDSHDSTFlagellin GVLY

RINTNINSMRTQEYMRQNQAKM TVADNRATLGATLNRLDFNVN N-SEQ ID NO: 150SNAMDRLSSGKRINNASDDAAGLAIATRM NLKSQSSAMAASASQIEDADM C-SEQ ID NO: 151RARESGLSVAADNTQNGMSLIRTADSAM AKEMSEMTKFKILNEAGISMLS Bacillus cereusNSVSNILLRMRDIANQSANGTNTDKNQVA QANQTPQMVSKLLQ strain Q1LQKEFAALKEQITYIADNTQFNDKNLLNGN [CDS of SEQ ID NO: 38] QTINIQTLDSHDSTFlagellin GFLN

RINTNINSMRTQEYMRQNQAKM LGATLNRLDFNVNNLKSQSSSM N-SEQ ID NO: 152SNAMDRLSSGKRINNASDDAAGLAIATRM ASAASQIEDADMAKEMSEMTK C-SEQ ID NO: 153RARESGLGVAANNTQDGMSLIRTADSALN FKILNEAGISMLSQANQTPQMBacillus thuringiensis SVSNILLRMRDIANQSANGTNTGDNQKAL VSKLLQserovar morrisoni DKEFSALKEQIDYISKNTEFNDKKLL [CDS of SEQ ID NO: 39]Flagellin GFLN

RINTNINSMRTQEYMRQNQTKM AIKSIDAALDTIASNRATLGATLN N-SEQ ID NO: 154SNAMDRLSSGKRINNASDDAAGLAIATRM RLDFNVNNLKSQSSSMASAAS C-SEQ ID NO: 155RARENGLGVAANNTQDGMSLIRTADSAL QIEDADMAKEMSEMTKFKILNBacillus thuringiensis NSVSNILLRMRDIANQSANGTNTSDNQKAEAGISMLSQANQTPQMVSKLL serovar neoleonensis LDKEFSALKEQIDYISKNTEENDKKLL Q[CDS of SEQ ID NO: 40] Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVNNLKSQS N-SEQ ID NO: 156SNAMDRLSSGKRINNASDDAAGLAIATRM SSMASAASQIEDADMAKEMSE C-SEQ ID NO: 157RARESGLGVAANNTQDGMSLIRTADSALN MTKFKILNEAGISMLSQANQTPBacillus thuringiensis SVSNILLRMRDIANQSANGTNTGDNQKAL QMVSKLLQserovar morrisoni DKEFSALKEQIDYISKNTEFNDKKLL [CDS of SEQ ID NO: 41]Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM RATLGATLNRLDFNVNNLKSQS N-SEQ ID NO: 158SNAMDRLSSGKRINNASDDAAGLAIATRM SSMASAASQIEDADMAKEMSE C-SEQ ID NO: 159RARESGLGVAANNTQDGMSLIRTADSALN MTKFKILNEAGISMLSQANQTPBacillus thuringiensis SVSNILLRMRDIANQSANGTNTGDNQKAL QMVSKLLQserovar morrisoni DKEFSALKEQIDYISKNTEFNDKKLL [CDS of SEQ ID NO: 42]Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM LGATLNRLDFNVNNLKSQQSS N-SEQ ID NO: 160SNAMDRLSSGKRINNASDDAAGLAIATRM MASAASQIEDADMAKEMSEM C-SEQ ID NO: 161RARESGLGVAANNTQDGMSLIRTADSAM TKFKILNEAGISMLSQANQTPQBacillus thuringiensis NSVSNILLRMRDIANQSANGTNTNGNQA MVSKLLQserovar jegathesan ALNKEFDALKQQINYISTNTEFNDKKLLDGS[CDS of SEQ ID NO: 43] NKTIAIQTLD Flagellin GVLN

RINTNINSMRTQEYMRQNQAKM DKIDEALKTIADNRATLGATLNR N-SEQ ID NO: 162SNAMDRLSSGKRINNASDDAAGLAIATRM LDFNVNNLKSQSASMASAASQI C-SEQ ID NO: 163RARESGLGVAANNTQDGMALIRTADSAM EDADMAKEMSEMTKFKILNEA Bacillus cereusNSVSNILLRRDIANQSANGTNTDKNQAAL GISMLSQANQTPQMVSKLLQ stain ATCC 10987QKEFGELQKQIDYIAGNTQFNDK [CDS of SEQ ID NO: 44] Flagellin WGFLI

RINTNINSMRTQEYMRQNQAK RATLGATLNRLDFNVNNLKSQQ N-SEQ ID NO: 164MSNAMDRLSSGKRINNASDDAAGLAIATR SSMASAASQIEDADMAKEMSE C-SEQ ID NO: 165MRARESGLGVAANNTQDGMSLIRTADSA MTKFKILNEAGISMLSQANQTPBacillus thuringiensis MNSVSNILLRMRDLANQSANGTNTNENQ QMVSKLLQserovar monterrey AALNKEFDALKEQINYISTNTEFNDKKLL [CDS of SEQ ID NO: 45]Flagellin WGFFY

RINTNINSMRTQEYMRQNQAK TVADNRATLGATLNRLDFNVN N-SEQ ID NO: 166MSNAMDRLSSGKRINNASDDAAGLAIATR NLKSQASSMAAAASQVEDAD C-SEQ ID NO: 167MRARESGLGVASNNTQDGMSLIRTADSAL MAKEMSEMTKFKILNEAGISM Bacillus cereusNSVSNILLRMRDLANQSANGTNTNENKAA LSQANQTPQMVSKLLQ strain NC7401MQKEFGELKEQIKYIAENTQFNDQHLL [CDS of SEQ ID NO: 46] Flagellin WGFFY

RINTNINSMRTQEYMRQNQAK TVADNRATLGATLNRLDFNVN N-SEQ ID NO: 168MSNAMDRLSSGKRINNASDDAAGLAIATR NLKSQASSMAAAASQVEDAD C-SEQ ID NO: 169MRARESGLGVASNNTQDGMSLIRTADSAL MAKEMSEMTKFKILNEAGISM Bacillus cereusNSVSNILLRMRDLANQSANGTNTNENKAA LSQANQTPQMVSKLLQ strain NC7401MQKEFGELKEQIKYIAENTQFNDQHLL [CDS of SEQ ID NO: 47] Flagellin (A-type)GVLN

RINTNINSLRTQEYMRQNQAKMS IDAALKTVADNRATLGATLNRL N-SEQ ID NO: 170NSMDRLSSGKRINNASDDAAGLAIATRMR DFNVNNLKSQSSSMASAASQIE C-SEQ ID NO: 171ARESGLNVAANNTQDGMSLIRTADSALGS DADMAKEMSEMTKFKILNEAG Bacillus cereusVSNILLRMRDLANQSANGTNTSDNQAAM ISMLSQANQTPQMVSKLLQ strain AH820QKEFAELQKQITYIADNTQFNDKNLL [CDS of SEQ ID NO: 48] Flagellin WGFFY

RINTNINSMRTQEYMRQNQAK TVADNRATLGATLNRLDFNVN N-SEQ ID NO: 172MSNAMDRLSSGKRINNASDDAAGLAIATR NLKSQASSMAAAASQVEDAD C-SEQ ID NO: 173MRARESGLGVASNNTQDGMSLIRTADSAL MAKEMSEMTKFKILNEAGISMBacillus cereus AH187 NSVSNILLRMRDLANQSANGTNTNENKAA LSQANQTPQMVSKLLQ[CDS of SEQ ID NO: 49] MQKEFGELKEQIKYIAENTQFNDQHLL Flagellin WGFFY

RINTNINSMRTQEYMRQNQAK TVADNRATLGATLNRLDFNVN N-SEQ ID NO: 174MSNAMDRLSSGKRINNASDDAAGLAIATR NLKSQASSAAAASQVEDADMA C-SEQ ID NO: 175MRARESGLGVASNNTQDGMSLIRTADSAL KEMSEMTKFKILNEAGISMLSQ Bacillus cereusNSVSNILLRMRDLANQSANGTNTNENKAA ANQTPQMVSKLLQ [CDS of SEQ ID NO: 50]MQKEFGELKEQIKYIAENTQFNDQHLL Flagellin protein Fla GFLN

RINTNINSMRTQEYMRQNQAKM LGATLNRLDFNVNNLKSQSSSM N-SEQ ID NO: 176SNAMDRLSSGKRINNASDDAAGLAIATRM ASAASQIEDADMAKEMSEMTK C-SEQ ID NO: 177RARESGLGVAANNTQDGMSLIRTADSALN FKILNEAGISMLSQANQTPQM Bacillus cereusSVSNILLRMRDIANQSANGTNTGDNQKAL VSKLLQ [CDS of SEQ ID NO: 51]DKEFSALKEQIDYISKNTEFNDKKLL Flagellin GFLN

RINTNINSMRTQEYMRQNQTKM RATLGATLNRLDFNVTNLKSQE N-SEQ ID NO: 178SNAMDRLSSGKRINNASDDAAGLAIATRM NSMAASASQIEDADMAKEMSE C-SEQ ID NO: 179RSREGGLNVAARNTEDGMSLIRTADSALN MTKFKILNEAGISMLSQANQTPBacillus thuringiensis SVSNILLRMRDLANQSASETNTSKNQAAM QMVSKLLQStrain HD-771 QKEFDQLKEQIQYI [CDS of SEQ ID NO: 52] Flagellin GFLN

RINTNINSMRTQEYMRQNQTKM RATLGATLNRLDFNVTNLKSQE N-SEQ ID NO: 180SNAMDRLSSGKRINNASDDAAGLAIATRM NSMAASASQIEDADMAKEMSE C-SEQ ID NO: 181RSREGGLNVAARNTEDGMSLIRTADSALN MTKFKILNEAGISMLSQANQTPBacillus thuringiensis SVSNILLRMRDLANQSASETNTSKNQAAM QMVSKLLQserovar sotto QKEFDQLKEQIQYI [CDS of SEQ ID NO: 53] Flagellin MGVLN

RINTNINSMRTQEYMRQNQAK AIKAIDEALETIASNRATLGATLN N-SEQ ID NO: 182MSTAMDRLSSGKRINNASDDAAGLAIATR RLDFNVNNLKNQASSMASAAS C-SEQ ID NO: 183MRARESGLGVAANNTQDGISLIRTADSAM QVEDADMAKEMSEMTKFKILNBacillus thuringiensis NSVSNILLRMRDLANQSANGTNTDKNQGEAGISMLSQANQTPQMVSKLL serovar Novosibirsk ALDKEFAALKEQIDYISKNTEFNDKKLL Q[CSD of SEQ ID NO: 54] Flagellin MGVLN

RINTNINSMRTQEYMRQNQAK AIDSALENIASNRATLGATLNRL N-SEQ ID NO: 184MSNAMDRLSSGKRINNASDDAAGLAIATR DFNVNNLKSQSSSMASAASQIE C-SEQ ID NO: 185MRARESGLGVAANNTQDGISLIRTADSAM DADMAKEMSEMTKFKILNEAGBacillus thuringiensis NSVSNILLRMRDLANQSANGTNTSENQAA ISMLSQANQTPQMVSKLLQserovar Londrina LDKEFGALKEQINYISTNTEFNDKKLL [CDS of SEQ ID NO: 55]Flagellin MGVLN

RINTNINSMRTQEYMRQNQAK LGATLNRLDFNVNNLKNQASS N-SEQ ID NO: 186MSTAMDRLSSGKRINNASDDAAGLAIATR MASAASQVEDADMAKEMSE C-SEQ ID NO: 187MRARESGLGVAANNTQDGISLIRTADSAM MTKFKILNEAGISMLSQANQTP Bacillus cereusNSVSNILLRMRDLANQSANGTNTDKNQG QMVSKLLQ strain E33LALDKEFAALKEQIDYISKNTEFNDKKLL [CDS of SEQ ID NO: 56] Flagellin MGVLN

RINTNINSMRTQEYMRQNQAK ATLNRLDFNVNNLKNQASSMA N-SEQ ID NO: 188MSTAMDRLSSGKRINNASDDAAGLAIATR SAASQVEDADMAKEMSEMTK C-SEQ ID NO: 189MRARESGLGVAANNTQDGISLIRTADSAM FKILNEAGISMLSQANQTPQM Bacillus cereusNSVSNILLRMRDLANQSANGTNTDKNQG VSKLLQ strain E33LALDKEFAALKEQIDYISKNTEFNDKKLL [CDS of SEQ ID NO: 57] Flagellin WGFFY

RINTNINSMRTQEYMRQNQAK AIAAIDAALTKVADNRATLGATL N-SEQ ID NO: 190MSTAMDRLSSGKRINNASDDAAGLAIATR NRLDFNVNNLKSQASSMASAA C-SEQ ID NO: 191MRARESGLGVAANNTQDGISLIRTADSAM SQVEDADMAKEMSEMTKFKIL Bacillus cereusNSVSNILLRMRDLANQSANGTNTDKNQA NEAGISMLSQANQTPQMVSKL strain FRI-35ALDKEFNALKEQIDYISKNTEFNDKKL LQ [CDS of SEQ ID NO: 58] Flagellin WGFFY

RIGTNVLSLNARQSLYENEKRM AIRKIEEALQNVSLHRADLGAMI N-SEQ ID NO: 192NVAMEHLATGKKLNNASDNPANIAIVTR NRLQFNIENLNSQSTALTDAAS C-SEQ ID NO: 193MHARASGMRVAIRNNEDAISMLRTAEAA RIEDADMAQEMSDFLKFKLLTE Bacillus cereusLQTVTNVLQRMRDLAVQSANGTNSNKNR VALSMVSQANQVPQMVSKLL strain FRI-35DSLNKEFQSLTEQIGYIDETTEFNN Q [CDS of SEQ ID NO: 59] Flagellin LVPFAVWLA

SRIRRRILDTDCKAESAVRIK MAASASQIEDADMAKEMSEM N-SEQ ID NO: 194EIPSDVLRAATERPLSCARIRVAIARPAASSE TKFKILSEAGISMLSQANQTPQ C-SEQ ID NO: 195ALLIRLPLDKRSIALLILAWFWRMYSCVRML MVSKLLQ Bacillus thuringiensisLMFVLILMLRTP [CDS of SEQ ID NO: 60] Flagellin AVWLA

SRIRRRILDTDCKAESAVRIKEIPS SMAASASQIEDADMAKEMSE N-SEQ ID NO: 196DVLRAATERPLSCARIRVAIARPAASSEALLI MTKFKILSEAGISMLSQANQTP C-SEQ ID NO: 197RLPLDKRSIALLILAWFWRMYSCVRMLLMF QMVSKLLQ Bacillus cereus VLILMLRTPstrain ATCC 4342 [CDS of SEQ ID NO: 61] Flagellin GFLN

RIGTNFLSMNARQSLYENEKRMN LGAMINRLHFNIENLNSQSMAL N-SEQ ID NO: 198VAMEHLATGKKLNHASDNPANIAIVTRMH TDAASRIEDADMAQEMSDFLK C-SEQ ID NO: 199ARANGMRVAIRNNEDAISMLRTAEAALQT FKLLTEVALSMVSQANQIPQMBacillus thuringiensis VMNILQRMRDLAIQSANSTNSNKNRDSLN VSKLLQ[CDS of SEQ ID NO: 62] KEFQSLTEQISYI Flagellin GFLN

RINTNINSMRTQEYMRQNQAKM LGATLNRLDFNVNNLKSQSSSM N-SEQ ID NO: 200SNAMDRLSSGKRINNASDDAAGLAIATRM ASAASQIEDADMAKEMSEMTK C-SEQ ID NO: 201RARESGLGVAANNTQDGMSLIRTADSALN FKILNEAGISMLSQANQTPQMBacillus thuringiensis SVSNILLRMRDIANQSANGTNTGDNQKAL VSKLLQ[CDS of SEQ ID NO: 63] DKEFSALKEQIDYI Flagellin

RINHNITALNTYRQFNNANNAQAKSME IDGAINQVSEQRSGLGATQNRL N-SEQ ID NO: 202KLSSGQRINSASDDAAGLAISEKMRGQIRG DHTINNLSTSSENLTASESRIRD C-SEQ ID NO: 203LDQASRNAQDGVSLIQTAEGALNETHDILQ VDYALAA Bacillus aryabhattaiRMRELVVQAGNGTNKTEDLDAIQDEIGSLI [CDS of SEQ ID NO: 64]EEIGGETDSKGISDRAQFNGRNLLDGSLDIT LQVGA Flagellin

RINTNINSMRTQEYMRQNQDKMNTSM IDQAIQDIADNRATYGSQLNRL N-SEQ ID NO: 204NRLSSGKQINSASDDAAGLAIATRMRAKE DHNLNNVNSQATNMAAAASQ C-SEQ ID NO: 205GGLNVGAKNTQDGMSALRTMDSALNSVS IEDADMAKEMSEMTKFKILSEABacillus manliponensis NILLRMRDLATQSATGTNQGNDRESLDLEGVSMLSQANQTPQMVSKLLQ [CDS of SEQ ID NO: 65]FQQLTEEITHIAEKTNFNGNALLSGSGSAIN VQLS Flagellin

RIGSWTATGMSIVNHMNRNWNAASKS LDEATKNVSMERSRLGAYQNRL N-SEQ ID NO: 206MLRLSSGYRINSAADDAAGLAISEKMRGQI EHAYNVAENTAINLQDAESRIR C-SEQ ID NO: 207RGLTMASKNIMDGVSLIQTAEGALNETHAI DVDIAKEMMNMVKSQILAQV Lysinibacillus sp.VQRMRELAVQAATDTNTDDDRAKLDLEF GQQVLAMHMQQAQGILRLLG strain BF-4QELKKEIDRISTDTEFNTRTLLNGDYKDNGL [CDS of SEQ ID NO: 66] KIQVG Flagellin

KIGSWTATGMSIVNHMNRNWNAASKS LDEATKNVSMERSRLGAYQNRL N-SEQ ID NO: 208MLRLSSGYRINSAADDAAGLAISEKMRGQI EHAYNVAENTAINLQDAESRIR C-SEQ ID NO: 209RGLTMASKNIMDGVSLIQTAEGALNETHAI DVDIAKEMMHMVKSQILAQV Lysinibacillus sp.VQRMRELAVQAATDTNTDDDRAKLDLEF GQQVLAMHIQQAQGILRLLG strain 13S34_airQELKKEIDRISTDTAFNTRTLLNGDYKDNGL [CDS of SEQ ID NO: 67] KIQVG Flagellin

IISHNLTALNTMNKLKQKDLAVSKSLGKL ISAAIDKVSAERARMGAYQNRL N-SEQ ID NO: 210SSGLRINGASDDAAGLAISEKMRGQIRGLN EHSRNNVVTYAENLTAAESRIR C-SEQ ID NO: 211QASRNIQDGISLIQVADGAMQEIHSMLQR DVDMAKEMMELMKNQIFTQA Paenibacillus sp.MNELAVQASNGTYSGSDRLNIQSEVEQLIE GQAMLLQTNTQPQAILQLLK strain HW567EIDEIAGNTGFNGIKLLNGNNEKTEKTEK [CDS of SEQ ID NO: 68] Flagellin

RINTNINSMRTQEYMRQNQAKMSNA IDSALETIASNRATLGATLNRLDF N-SEQ ID NO: 212MDRLSSGKRINNASDDAAGLAIATRMRAR NVNNLKSQSSAMASAASQIED C-SEQ ID NO: 213ESGLGVAANNTQDGMSLIRTADSAMNSV ADMAKEMSEMTKFKILNEAGIS Bacillus anthracisSNILLRMRDLANQSANGTNTKENQDALDK MLSQANQTPQMVSKLLQ [CDS of SEQ ID NO: 69]EFGALKEQIDYISKNTEFNDKKLLNGDNKSI AIQTL Flagellin

QKSQYKKMGVLKMRINTNINSMRTQEY ALNTVAGNRATLGATLNRLDR N-SEQ ID NO: 214MRQNQDKMNVSMNRLSSGKRINSAADD NVENLNNQATNMASAASQIED C-SEQ ID NO: 215AAGLAIATRMRARQSGLEKASQNTQDGM ADMAKEMSEMTKFKILNEAGIS Bacillus anthracisSLIRTAESAMNSVSNILTRMRDIAVQSSNG MLSQANQTPQMVSKLLQ [CDS of SEQ ID NO: 70]TNTAENQSALQKEFAELQEQIDYIAKNTEF NDKNLLAGTGAVTIGSTSISGAEISIETL Flagellin

RINTNINSMRTQEYMRQNQDKMNVS ALNTVAGNRATLGATLNRLDR N-SEQ ID NO: 216MNRLSSGKRINSAADDAAGLAIATRMRAR NVENLNNQATNMASAASQIKD C-SEQ ID NO: 217QSGLEKASQNTQDGMSLIRTAESAMNSVS ADKAKEMSEMTKFKILNEAGIS Bacillus anthracisNILTRMRDIAVQSSNGTNTAENQSALQKE MLSQANQTPQMVSKLLQ [CDS of SEQ ID NO: 71]FAELQEQIDYIAKNTEFNDKNLLAGTGAVTI GSTSISGAEISIETL Flagellin

RINTNINSMRTQEYMRQNQDKMNVS ALNTVAGNRATLGATLNRLDR N-SEQ ID NO: 218MNRLSSGKRINSAADDAAGLAIATRMRAR NVENLNNQATNMASAASQIED C-SEQ ID NO: 219QSGLEKASQNTQDGMSLIRTAESAMNSVS ADMAKEMSEMTKFKILNEAGIS Bacillus anthracisNILTRMRDIAVQSSNGTNTAENQSALQKE MLSQANQTPQMV [CDS of SEQ ID NO: 72]FAELQEQIDYIAKNTEFNDKNLLAGTGAVTI GSTSISGAEISIETL Flagellin

NVSMNRLSSGKRINSAADDAAGLAIATR LNTALNTVAGNRATLGATLNRL N-SEQ ID NO: 220MRARQSGLEKASQNTQDGMSLIRTAESA DRNVENLNNQATNMASAASQI C-SEQ ID NO: 221MNSVSNILTRMRDIAVQSSNGTNTAENQS EDADMAKEMSEMTKFKILNEA Bacillus anthracisALQKEFAELQEQIDYIAKNTEFNDKNLLAG GISMLSQANQTPQMVSKLLQ strain H9401TGAVTIGSTSISGAEISIETL [CDS of SEQ ID NO: 73] Flagellin

RINHNITALNTYRQFNNANNAQAKSME IIDGAINQVSEQRSGLGATQNR N-SEQ ID NO: 222KLSSGQRINSASDDAAGLAISEKMRGQIRG LDHTINNLSTSSENLTASESRIRD C-SEQ ID NO: 223LDQASRNAQDGVSLIQTAEGALNETHDILQ VDYALAA Bacillus megateriumRMRELVVQAGNGTNKTEDLDAIQDEIGSLI strain WSH-002EEIGGEADSKGISDRAQFNGRNLLDGSLDIT [CDS of SEQ ID NO: 74] LQVGA Flagellin

RINHNLPALNAYRNLAQNQIGTSKILERL FKAAIDQVSRIRSYFGAIQNRLE N-SEQ ID NO: 224SSGYRINRASDDAAGLAISEKMRGQIRGLE HVVNNLSNYTENLTGAESRIRD C-SEQ ID NO: 225QGQRNTMDGVSLIQTAEGALQEIHEMLQ ADMAKEMTEFTRFNIINQSATA AneurinibacillusRMRELAVQAANGTYSDKDKKAIEDEINQL MLAQANQLPQGVLQLLKG sp. XH2TAQIDQIAKTTEFNGIQLIGDSDSTSLQDVK [CDS of SEQ ID NO: 75]

The amino acid sequence of the flagellin or flagellin-associatedpolypeptide can comprise any one of SEQ ID NOs: 226-300, or anycombination thereof.

The amino acid sequence of the flagellin or flagellin-associatedpolypeptide can comprise SEQ ID NO: 226.

The amino acid sequence of the flagellin or flagellin-associatedpolypeptide can comprise any one of SEQ ID NOs: 301-375, or anycombination thereof.

The amino acid sequence of the flagellin or flagellin-associatedpolypeptide can comprise SEQ ID NO: 301.

The flagellin-derived polypeptide sequence for Bt4Q7Flg22 (SEQ ID NO:226) was identified from a proprietary “in house” library from Bacillusthuringiensis (Bt.) strain 4Q7. Conserved primers to full lengthflagellin from E. coli were used to screen the Bt.4Q7 strain library andidentify a functional flagellin-associated bioactive priming Flg22polypeptide.

TABLE 3 Flagellin polypeptides Flg22 and FlgII-28identified from Bacillus spp. SEQ ID NO: Peptide Flg22 Flg22-Bt.4Q7DRLSSGKRINSASDDAAGLAIA SEQ ID NO: 226 Bacillus  thuringiensis strain 4Q7Flg22 DRLSSGKRINSASDDAAGLAIA SEQ ID NO: 227 Bacillus  thuringiensis,strain HD1002 Flg22 DRLSSGKRINSASDDAAGLAIA SEQ ID NO: 228 Bacillus thuringiensis, strain HD-789 Flg22 DRLSSGKRINSASDDAAGLAIA SEQ ID NO: 229Bacillus cereus strain G9842 Flg22 EHLATGKKLNNASDNPANIAIV SEQ ID NO: 230Bacillus  thuringiensis serovar indiana strain HD521 Flg22DRLSSGKRINNASDDAAGLAIAT SEQ ID NO: 231 Bacillus  thuringiensisstrain CTC Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 232 Bacillusthuringiensis serovar  yunnanensis strain IEBC- T20001 Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 233 Bacillus  thuringiensis serovar tolworthi Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 234 Bacillus cereusstrain FM1 Flg22 EHLATGKKLNHASDNPANVAIV SEQ ID NO: 235 Bacillus cereusstrain FM1 Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 236 Bacillus thuringiensis strain MC28 Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 237Bacillus  bombysepticus strain Wang Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 238 Bacillus  thuringiensis serovar kenyae Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 239 Bacillus  thuringiensisserovar kenyae Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 240Bacillus cereus Flg22 EHLATGKKLNNASDNPANIAIV SEQ ID NO: 241Bacillus cereus Flg22 EHLATGKKLNHASDNPANVAIV SEQ ID NO: 242 Bacillus thuringiensis serovar finitimus strain YBT-020 Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 243 Bacillus  thuringiensisserovar finitimus strain YBT-020 Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 244 Bacillus cereus stain B4264 Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 245 Bacillus  thuringiensis serovar  nigeriensis Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 246 Bacillus  thuringiensis Flg22EHFATGKKLNHASDNPANVAIV SEQ ID NO: 247 Bacillus  thuringiensisserovar konkukian strain 97-27 Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 248 Bacillus  thuringiensis serovar konkukian strain 97-27Flg22 EHLATGKKLNHASDNPANIVIV SEQ ID NO: 249 Bacillus  thuringiensisserovar  thuringiensis strain IS5056 Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 250 Bacillus  thuringiensis serovar  thuringiensisstrain IS5056 Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 251 Bacillus thuringiensis strain Bt407 Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 252Bacillus  thuringiensis serovar chinensis CT-43 Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 253 Bacillus  thuringiensisserovar canadensis Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 254 Bacillus thuringiensis serovar galleriae Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 255 Bacillus weihenstephanensis Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 256 Bacillus  thuringiensis serovar ostriniae Flg22EHLATGKKLNHASDNPANVAIV SEQ ID NO: 257 Bacillus  thuringiensis Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 258 Bacillus  thuringiensis Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 259 Bacillus  thuringiensis serovar pondicheriensis Flg22 EHLATGKKLNHASDNPANIVIV SEQ ID NO: 260 Bacillus thuringiensis serovar Berliner Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 261 Bacillus  thuringiensis serovar Berliner Flg22EHLATGKKLNHASNNPANVAIV SEQ ID NO: 262 Bacillus cereus strain Q1 Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 263 Bacillus cereus strain Q1 Flg22EHLATGKKLNHASDNPANIAIV SEQ ID NO: 264 Bacillus  thuringiensisserovar morrisoni Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 265 Bacillus thuringiensis serovar  neoleonensis Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 266 Bacillus  thuringiensis serovar morrisoni Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 267 Bacillus  thuringiensisserovar morrisoni Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 268 Bacillus thuringiensis serovar jegathesan Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 269 Bacillus cereus stain ATCC 10987 Flg22 from DRLSSGKRINNASDDAAGLAIA Flagellin A SEQ ID NO: 270 Bacillus thuringiensis serovar monterrey Flg22 EHLATGKKLNNASDNPANIAIVSEQ ID NO: 271 Bacillus cereus strain NC7401 Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 272 Bacillus cereus strain NC7401Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 273 Bacillus cereus strain AH820Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 274 Bacillus cereus AH187 Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 275 Bacillus cereus Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 276 Bacillus cereus Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 277 Bacillus  thuringiensisStrain HD-771 [51] Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 278 Bacillus thuringiensis serovar sotto [52] Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 279 Bacillus  thuringiensis serovar  Novosibirsk Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 280 Bacillus  thuringiensisserovar londrina Flg22 EHLATGKKLNHASNNPANIAIV SEQ ID NO: 281Bacillus cereus strain E33L Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 282Bacillus cereus strain E33L Flg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 283Bacillus cereus strain FRI-35 Flg22 EHLATGKKLNNASDNPANIAIVSEQ ID NO: 284 Bacillus cereus strain FRI-35 Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 285 Bacillus  thuringiensis Flg22DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 286 Bacillus cereus strain ATCC 4342Flg22 EHLATGKKLNHASDNPANIAIV SEQ ID NO: 287 Bacillus  thuringiensisFlg22 DRLSSGKRINNASDDAAGLAIA SEQ ID NO: 288 Bacillus  thuringiensisFlg22 EKLSSGQRINSASDDAAGLAIS SEQ ID NO: 289 Bacillus  aryabhattai Flg22NRLSSGKQINSASDDAAGLAIA SEQ ID NO: 290 Bacillus  manliponensis Flg22LRLSSGYRINSAADDAAGLAIS SEQ ID NO: 291 Lysinibacillus sp. strain BF-4Flg22 LRLSSGYRINSAADDAAGLAIS SEQ ID NO: 292 Lysinibacillus sp.strain 13S34_air Flg22 GKLSSGLRINGASDDAAGLAIS SEQ ID NO: 293Paenibacillus sp. strain HW567 Flg22 DRLSSGKRINNASDDAAGLAIASEQ ID NO: 294 Bacillus  anthracis Flg22 NRLSSGKRINSAADDAAGLAIASEQ ID NO: 295 Bacillus  anthracis Flg22 NRLSSGKRINSAADDAAGLAIASEQ ID NO: 296 Bacillus  anthracis Flg22 NRLSSGKRINSAADDAAGLAIASEQ ID NO: 297 Bacillus  anthracis Flg22 NRLSSGKRINSAADDAAGLAIASEQ ID NO: 298 Bacillus  anthracis strain H9401 Flg22EKLSSGQRINSASDDAAGLAIS SEQ ID NO: 299 Bacillus  megateriumstrain WSH-002 Flg22 ERLSSGYRINRASDDAAGLAIS SEQ ID NO: 300Aneurinibacillus sp. XH2 Peptide Flg15 Flg15-Bt4Q7 RINSAKDDAAGLAIASEQ ID NO: 752 Modified FLG15- Bt4Q7; Syn01 Bacillus  thuringiensisstrain 4Q7 Peptide FglI-28 FlgII-28-Bt.4Q7 SVSNILLRMRDLANQSANGTNTKGNQASSEQ ID NO: 301 Bacillus  thuringiensis strain 4Q7 FlgII-28SVSNILLRMRDLANQSANGTNTKGNQAS SEQ ID NO: 302 Bacillus  thuringiensis,strain HD1002 FlgII-28 SVSNILLRMRDLANQSANGTNTKGNQAS SEQ ID NO: 303Bacillus  thuringiensis, strain HD-789 FlgII-28SVSNILLRMRDLANQSANGTNTKGNQAS SEQ ID NO: 304 Bacillus cereus strain G9842FlgII-28 TVTNILQRMRDLAVQSANGTNSNKNRHS SEQ ID NO: 305 Bacillus thuringiensis serovar indiana strain HD521 FlgII-28SVSNILLRMRDIANQSANITNTNENKSA SEQ ID NO: 306 Bacillus  thuringiensisstrain CTC FlgII-28 SVSNILLRMRDLANQSANGTNTDDNQKA SEQ ID NO: 307 Bacillusthuringiensis serovar  yunnanensis strain IEBC- T20001 FlgII-28SVSNILLRMRDLANQSANGTNTDENKAA SEQ ID NO: 308 Bacillus  thuringiensisserovar tolworthi FlgII-28 SVSNILLRMRDIANQSANGTNTDKNQVA SEQ ID NO: 309Bacillus cereus strain FM1 FlgII-28 TVTNILQRMRDVAVQSANGTNSNKNRDSSEQ ID NO: 310 Bacillus cereus strain FM1 FlgII-28SVSNILLRMRDIANQSANGTNTADNQQA SEQ ID NO: 311 Bacillus  thuringiensisstrain MC28 FlgII-28 SVSNILLRMRDLANQSASGTNTDKNQAA SEQ ID NO: 312Bacillus  bombysepticus strain Wang FlgII-28SVSNILLRMRDLANQSASGTNTDKNQAA SEQ ID NO: 313 Bacillus  thuringiensisserovar kenyae FlgII-28 SVSNILLRMRDLANQSASGTNTDKNQAA SEQ ID NO: 314Bacillus  thuringiensis serovar kenyae FlgII-28SVSNILLRMRDLANQSANGTNTGDNQKA SEQ ID NO: 315 Bacillus cereus FlgII-28TNILQRMRDLAVQSANGTNSNKNRDSLN SEQ ID NO: 316 Bacillus cereus FlgII-28TNVLQRMRDVAVQSANGTNLNKNRDSLN SEQ ID NO: 317 Bacillus  thuringiensisserovar finitimus strain YBT-020 FlgII-28 SVSNILLRMRDIANQSANGTNTDSNKSASEQ ID NO: 318 Bacillus  thuringiensis serovar finitimus strain YBT-020FlgII-28 SVSNILLRMRDLANQSANGTNTAENKAA SEQ ID NO: 319 Bacillus cereusstain B4264 FlgII-28 SVSNILLRMRDIANQSANGTNTSDNQKA SEQ ID NO: 320Bacillus  thuringiensis serovar  nigeriensis FlgII-28SVSNILLRMRDIANQSANGTNTADNQQA SEQ ID NO: 321 Bacillus  thuringiensisFlgII-28 TVMNILQRMRDLAVQSANGTNSNKNRDS SEQ ID NO: 322 Bacillusthuringiensis serovar konkukian strain 97-27 FlgII-28SVSNILLRMRDIANQSANGTNTADNQQA SEQ ID NO: 323 Bacillus  thuringiensisserovar konkukian strain 97-27 FlgII-28 TVTNILQHMRDFAIQSANGTNSNTNRDSSEQ ID NO: 324 Bacillus  thuringiensis serovar  thuringiensisstrain IS5056 FlgII-28 SVSNILLRMRDISNQSANGTNTDKNQSA SEQ ID NO: 325Bacillus  thuringiensis serovar  thuringiensis strain IS5056 FlgII-28SVSNILLRMRDISNQSANGTNTDKNQSA SEQ ID NO: 326 Bacillus  thuringiensisstrain Bt407 FlgII-28 SVSNILLRMRDISNQSANGTNTDKNQSA SEQ ID NO: 327Bacillus  thuringiensis serovar chinensis CT-43 FlgII-28SVSNILLRMRDLANQSANGTNTNENQAA SEQ ID NO: 328 Bacillus  thuringiensisserovar canadensis FlgII-28 SVSNILLRMRDLANQSANGTNTNENQAA SEQ ID NO: 329Bacillus  thuringiensis serovar galleriae FlgII-28SVSNILLRMRDLSNQSANGTNTDENQQA SEQ ID NO: 330 Bacillus weihenstephanensisFlgII-28 SVSNILLRMRDIANQSANGTNTGDNQKA SEQ ID NO: 331 Bacillus thuringiensis serovar ostriniae FlgII-28 TVANILQRMRDLAVQSSNDTNSNKNRDSSEQ ID NO: 332 Bacillus  thuringiensis FlgII-28SVSNILLRMRDLANQSANGTNTDDNQKA SEQ ID NO: 333 Bacillus  thuringiensisFlgII-28 SVSNILLRMRDLANQSANGTNTDDNQKA SEQ ID NO: 334 Bacillus thuringiensis serovar  pondicheriensis FlgII-28TVTNILQHMRDFAIQSANGTNSNTNRDS SEQ ID NO: 335 Bacillus  thuringiensisserovar Berliner FlgII-28 SVSNILLRMRDISNQSANGTNTDKNQSA SEQ ID NO: 336Bacillus  thuringiensis serovar Berliner FlgII-28TVTNVLQRMRDVAVQSANGTNSSKNRDS SEQ ID NO: 337 Bacillus cereus strain Q1FlgII-28 SVSNILLRMRDIANQSANGTNTDKNQVA SEQ ID NO: 338 Bacillus cereusstrain Q1 FlgII-28 TVMNILQRMRDLAIQSANSTNSNKNRDS SEQ ID NO: 339 Bacillus thuringiensis serovar morrisoni FlgII-28 SVSNILLRMRDIANQSANGTNTSDNQKASEQ ID NO: 340 Bacillus  thuringiensis serovar  neoleonensis FlgII-28SVSNILLRMRDIANQSANGTNTGDNQKA SEQ ID NO: 341 Bacillus  thuringiensisserovar morrisoni FlgII-28 SVSNILLRMRDIANQSANGTNTGDNQKA SEQ ID NO: 342Bacillus  thuringiensis serovar morrisoni FlgII-28SVSNILLRMRDIANQSANGTNTNGNQAA SEQ ID NO: 343 Bacillus  thuringiensisserovar jegathesan FlgII-28 SVSNILLRMRDIANQSANGTNTDKNQAA SEQ ID NO: 344Bacillus cereus stain ATCC 10987 FlgII-28 fromSVSNILLRMRDLANQSANGTNTNENQAA Flagellin A SEQ ID NO: 345 Bacillus thuringiensis serovar monterrey FlgII-28 TVTNVLQRMRDLAVQSANDTNSNKNRDSSEQ ID NO: 346 Bacillus cereus strain NC7401 FlgII-28SVSNILLRMRDLANQSANGTNTNENKAA SEQ ID NO: 347 Bacillus cereusstrain NC7401 FlgII-28 SVSNILLRMRDLANQSANGTNTSDNQAA SEQ ID NO: 348Bacillus cereus strain AH820 FlgII-28 SVSNILLRMRDLANQSANGTNTNENKAASEQ ID NO: 349 Bacillus cereus AH187 FlgII-28SVSNILLRMRDLANQSANGTNTNENKAA SEQ ID NO: 350 Bacillus cereus FlgII-28SVSNILLRMRDIANQSANGTNTGDNQKA SEQ ID NO: 351 Bacillus cereus FlgII-28SVSNILLRMRDLANQSASETNTSKNQAA SEQ ID NO: 352 Bacillus  thuringiensisStrain HD-771 [51] FlgII-28 SVSNILLRMRDLANQSASETNTSKNQAA SEQ ID NO: 353Bacillus  thuringiensis serovar sotto [52] FlgII-28SVSNILLRMRDIANQSANGTNTGDNQKA SEQ ID NO: 354 Bacillus  thuringiensisserovar  Novosibirsk FlgII-28 SVSNILLRMRDLANQSANGTNTSENQAASEQ ID NO: 355 Bacillus  thuringiensis serovar londrina FlgII-28TVTNILQRMRDLAVQSANVTNSNKNRNS SEQ ID NO: 356 Bacillus cereus strain E33LFlgII-28 SVSNILLRMRDLANQSANGTNTDKNQGA SEQ ID NO: 357 Bacillus cereusstrain E33L FlgII-28 SVSNILLRMRDLANQSANGTNTDKNQAA SEQ ID NO: 358Bacillus cereus strain FRI-35 FlgII-28 TVTNVLQRMRDLAVQSANGTNSNKNRDSSEQ ID NO: 359 Bacillus cereus strain FRI-35 FlgII-28SVSNILLRMRDIANQTANGTNKDTDIEA SEQ ID NO: 360 Bacillus  thuringiensisFlgII-28 SVSNILLRMRDIANQTANGTNKDTDIEA SEQ ID NO: 361 Bacillus cereusstrain ATCC 4342 FlgII-28 TVMNILQRMRDLAIQSANSTNSNKNRDS SEQ ID NO: 362Bacillus  thuringiensis FlgII-28 SVSNILLRMRDIANQSANGTNTGDNQKASEQ ID NO: 363 Bacillus  thuringiensis FlgII-28ETHDILQRMRELVVQAGNGTNKTEDLDA SEQ ID NO: 364 Bacillus  aryabhattaiFlgII-28 SVSNILLRMRDLATQSATGTNQGNDRES SEQ ID NO: 365 Bacillus manliponensis FlgII-28 ETHAIVQRMRELAVQAATDTNTDDDRAK SEQ ID NO: 366Lysinibacillus sp. strain BF-4 FlgII-28 ETHAIVQRMRELAVQAATDTNTDDDRAKSEQ ID NO: 367 Lysinibacillus sp. strain 13S34_air FlgII-28EIHSMLQRMNELAVQASNGTYSGSDRLN SEQ ID NO: 368 Paenibacillus sp.strain HW567 FlgII-28 SVSNILLRMRDLANQSANGTNTKENQDA SEQ ID NO: 369Bacillus anthracis FlgII-28 SVSNILTRMRDIAVQSSNGTNTAENQSA SEQ ID NO: 370Bacillus anthracis FlgII-28 SVSNILTRMRDIAVQSSNGTNTAENQSA SEQ ID NO: 371Bacillus anthracis FlgII-28 SVSNILTRMRDIAVQSSNGTNTAENQSA SEQ ID NO: 372Bacillus anthracis FlgII-28 SVSNILTRMRDIAVQSSNGTNTAENQSA SEQ ID NO: 373Bacillus anthracis strain H9401 FlgII-28 ETHDILQRMRELVVQAGNGTNKTEDLDASEQ ID NO: 374 Bacillus  megaterium strain WSH-002 FlgII-28EIHEMLQRMRELAVQAANGTYSDKDKKA SEQ ID NO: 375 Aneurinibacillus sp. XH2

Retro-Inverso Flagellin-Associated Polypeptides

Bioactive Flg polypeptide(s) useful for priming can be created in anon-natural isomeric or retro-inverso (RI) form.

The retro-inverso Flg polypeptides can exhibit enhanced binding affinityfor the FLS receptor protein(s). Plant flagellin receptors, like FLS2,can recognize a retro inverso Flg polypeptide fragment such as eitherFlg22 or FlgII-28 located within the N-terminal conserved domain offlagellin. The retro-inverso forms of these Flg polypeptides areprovided as biologically active forms, which can recognize and interactwith the Flg-associated or FLS receptor protein on the surface of theplant cell membrane.

Retro-inverso Flg polypeptides can possess an increased activity andstability to proteolytic degradation at the plant membrane surface. Forexample, retro inverso forms of Bacillus Flg22 or FlgII-28 polypeptidescan increase activity and stability of the Flg polypeptide(s) andincrease protection against proteolytic degradation at the plant surfaceor root surface. The retro inverso forms also exhibit enhanced stabilitywhen applied in a field, or on or in a soil.

Retro-inverso polypeptides are topological mirror images of the nativestructures of the parent polypeptide. Retro inverso synthetic forms ofthe polypeptide sequences are created by reversing the polypeptidesequences and using retro-all-D or retro-enantio-peptides. The allD-chain amino acid Flg polypeptide(s) adopts a “mirror image” of thethree-dimensional structure of its related L-peptide or L-chain amino.

This is further accomplished by creating a retro-inverso alteration ofany of the parent Flg polypeptide derived from Bacillus or otherEubacteria in Table 3. Retro-inverso polypeptides that were designed tothe Flg22 (RI Flg22: SEQ ID NOs: 376-450), and FlgII-28 (RI-FlgII-28:SEQ ID NOs: 451-525) are provided in Table 4. Retro inverso forms ofEc.Flg22 (SEQ ID NO: 526) and EcFlg15 (SEQ ID NO: 529) as provided inTable 5 were also created from E. coli derived sequences.

The polypeptide can include a retro inverso Flg22 polypeptide.

The polypeptide can comprise a retro inverso FlgII-28 polypeptide.

Any of the flagellin-associated bioactive priming polypeptidescomprising Bacillus or from other Eubacteria Flg22 or FlgII-28polypeptides in Table 3 can be used in their retro-inversed forms(referenced in Table 4).

Retro inverso forms of the Flg bioactive priming polypeptides asreferenced herein can be provided in any of three forms where theinversion of amino acid chirality contains the normal-all-D (inverso),all-L (retro) and/or retro-all-D (retro-inverso) or a combination ofthese forms to achieve the desired phenotypes in a plant.

The Bacillus-derived L-Flg22 and L-FlgII-28 polypeptides in Table 3 andthe E.c. native L-Flg22 and L-Flg15 polypeptides in Table 5 weresynthetically generated via retro-inverso engineering to formretro-inverso D-Flg22 polypeptide (SEQ ID NO: 376-450), D-FlgII-28 (SEQID NO: 451-525), and E.c. D-Flg22 polypeptide (SEQ ID NO: 527, 529).

The inversion of amino acid chirality (all-L to all-D) for Bt.4Q7 Flg22(SEQ ID NO: 376), which is provided as a small linear polypeptidefragment and is referred to as a retro inverso modification was achievedby a reversal of the direction of the polypeptide backbone and describedbelow.

-   -   (^(D)A^(D)IA^(D)L^(D)G^(D)A^(D)A^(D)D^(D)D^(D)S^(D)A^(D)S^(D)N^(D)I^(D)R^(D)K^(D)G^(D)S^(D)S^(D)L^(D)R^(D)D)

The retro inverso all D-chain amino acid Flg22 polypeptide adopts a“mirror image” of the three-dimensional structure of its related nativeL-Bt.4Q7Flg 22 polypeptide and this all L-chain has an equivalent mirrorimage to the all D Bt.4Q7Flg22 polypeptide. All L-amino acid residuesare replaced by their D-enantiomers leading to all D-peptides or retroall D-isomer-peptides containing amide linkages. The native L-amino acidchain form of Bt.4Q7 Flg22 polypeptide chain reversed to generate theretro-inverso synthetic all-D confirmation that is prepared by replacingall the L-amino acid residues with their corresponding D-enantiomers.

FIG. 1 provides a diagrammatic representation of a natural (all L)Bt.4Q7 Flg22 and its retro inverso or mirror image to form an all DBt.4Q7 Flg22 enantiomeric polypeptide. The retro-inverso Flg polypeptidethat corresponds to Bt.4Q7 Flg22 (SEQ ID NO: 226) is described as SEQ IDNO: 376.

In the case of short polypeptides, such as Flg22, Flg15 and FlgII-28,the mirroring of the side chain positions in a conformational changefrom L-to-D conversion states results in a mirroring of symmetrytransformations of the side chains as well.

Retro-all-D analogues have been found to possess biological activity(Guptasarma, “Reversal of peptide backbone direction may result inmirroring of protein structure, FEBS Letters 310: 205-210, 1992). Theretro-inverso D-Flg polypeptide(s) can assume a side chain topology inits extended conformation that is similar to a corresponding nativeL-Flg polypeptide sequence, thus emulating biological activities of thenative L-parent molecule while fully resistant to proteolyticdegradation thus increasing stability when the polypeptide contacts theplant or the surrounding environment.

Retro-inverso Flg bioactive priming polypeptides are described in Table4 or Table 5. Retro inverso Flg-associated bioactive primingpolypeptides provided in Table 4 were selected for their enhancedactivity and stability and their ability to survive under varyingconditions and environments. Based on their D enantiomer nature, theyare more resistant to proteolytic degradation and can survive and existin harsher environmental conditions.

TABLE 4 Retro-inverso flagellin polypeptides from Flg22and FlgII-28 from Bacillus SEQ ID NO: Peptide Flg22 RI Bt.4Q7Flg22AIALGAADDSASNIRKGSSLRD SEQ ID NO: 376 Bacillus  thuringiensis strain 4Q7RI Flg22 AIALGAADDSASNIRKGSSLRD SEQ ID NO: 377 Bacillus  thuringiensis,strain HD1002 RI Flg22 AIALGAADDASNIRKGSSLRD SEQ ID NO: 378 Bacillus thuringiensis, strain HD-789 RI Flg22 AIALGAADDSASNIRKGSSLRDSEQ ID NO: 379 Bacillus cereus strain G9842 RI Flg22VIANAPNDSANNLKKGTALHE SEQ ID NO: 380 Bacillus  thuringiensisserovar indiana strain HD521 RI Flg22 TAIAGAADDSANNIRKGSSLRDSEQ ID NO: 381 Bacillus  thuringiensis strain CTC RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 382 Bacillus  thuringiensisserovaryunnanensis strain IEBC-T20001 RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 383 Bacillus  thuringiensis serovar tolworthi RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 384 Bacillus cereus strain FM1RI Flg22 VIAVNAPNDSAHNLKKGTALHE SEQ ID NO: 385 Bacillus cereusstrain FM1 RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 386 Bacillus thuringiensis strain MC28 RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 387Bacillus  bombysepticus strain Wang RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 388 Bacillus  thuringiensis serovar kenyae RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 389 Bacillus  thuringiensisserovar kenyae RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 390Bacillus cereus RI Flg22 VIAINAPNDASNNLKKGTALHE SEQ ID NO: 391Bacillus cereus RI Flg22 VIANAPNDSAHNLKKGTALHE SEQ ID NO: 392 Bacillus thuringiensis serovar finitimus strain YBT-020 RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 393 Bacillus  thuringiensisserovar finitimus strain YBT-020 RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 394 Bacillus cereus stain B4264 RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 395 Bacillus  thuringiensis serovar nigeriensis RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 396 Bacillus thuringiensis RI Flg22 VIANAPNDSAHNLKKGTAFHE SEQ ID NO: 397 Bacillus thuringiensis serovar konkukian strain 97-27 RI Flg22AIALGAADDSANNRKGSSLRD SEQ ID NO: 398 Bacillus  thuringiensisserovar konkukian strain 97-27 RI Flg22 VIVINAPNDSAHNLKKGTALHESEQ ID NO: 399 Bacillus  thuringiensis serovar  thuringiensisstrain IS5056 RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 400 Bacillus thuringiensis serovar  thuringiensis strain IS5056 RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 401 Bacillus  thuringiensisstrain Bt407 RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 402 Bacillus thuringiensis serovar chinensis CT-43 RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 403 Bacillus  thuringiensis serovar canadensis RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 404 Bacillus  thuringiensisserovar galleriae RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 405Bacillus weihenstephanensis RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 406 Bacillus  thuringiensis serovar ostriniae RI Flg22VIANAPNDSAHNLKKGTALHE SEQ ID NO: 407 Bacillus  thuringiensis RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 408 Bacillus  thuringiensis RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 409 Bacillus  thuringiensis serovar pondicheriensis RI Flg22 VIVINAPNDASHNLKKGTALHE SEQ ID NO: 410 Bacillus thuringiensis serovar Berliner RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 411 Bacillus  thuringiensis serovar Berliner RI Flg22VIAVANPNNSAHNLKKGTALHE SEQ ID NO: 412 Bacillus cereus strain Q1 RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 413 Bacillus cereus strain Q1 RI Flg22VIANAPNDSAHNLKKGTALHE SEQ ID NO: 414 Bacillus  thuringiensisserovar morrisoni RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 415Bacillus  thuringiensis serovar  neoleonensis RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 416 Bacillus  thuringiensisserovar morrisoni RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 417Bacillus  thuringiensis serovar morrisoni RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 418 Bacillus  thuringiensisserovar jegathesan RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 419Bacillus cereus stain ATCC 10987 AIALGAADDASNNIRKGSSLRD RI Flg22 fromFlagellin A SEQ ID NO: 420 Bacillus  thuringiensis serovar monterreyRI Flg22 VIANAPNDSANNLKKGTALHE SEQ ID NO: 421 Bacillus cereusstrain NC7401 RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 422Bacillus cereus strain NC7401 RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 423 Bacillus cereus strain AH820 RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 424 Bacillus cereus AH187 RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 425 Bacillus cereus RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 426 Bacillus cereus RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 427 Bacillus  thuringiensisStrain HD-771 [51] RI Flg22 AIALGAADDANNIRKGSSLRD SEQ ID NO: 428Bacillus  thuringiensis serovar sotto [52] RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 429 Bacillus  thuringiensis serovar Novosibirsk RI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 430 Bacillus thuringiensis serovar londrina RI Flg22 VIAINAPNNSAHNLKKGTALHESEQ ID NO: 431 Bacillus cereus strain E33L RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 432 Bacillus cereus strain E33LRI Flg22 AIALGAADDSANNIRKGSSLRD SEQ ID NO: 433 Bacillus cereusstrain FRI-35 RI Flg22 VIAINAPNDSANNLKKGTALHE SEQ ID NO: 434Bacillus cereus strain FRI-35 RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 435 Bacillus  thuringiensis RI Flg22 AIALGAADDANNIRKGSSLRDSEQ ID NO: 436 Bacillus cereus strain ATCC 4342 RI Flg22VIANAPNDSAHNLKKGTALHE SEQ ID NO: 437 Bacillus  thuringiensis RI Flg22AIALGAADDSANNIRKGSSLRD SEQ ID NO: 438 Bacillus  thuringiensis RI Flg22SIALGAADDSASNIRQGSSLKE SEQ ID NO: 439 Bacillus  aryabhattai RI Flg22AIALGAADDSASNIQKGSSLRN SEQ ID NO: 440 Bacillus  manliponensis RI Flg22SIALGAADDAASNIRYGSSLRL SEQ ID NO: 441 Lysinibacillus sp. strain BF-4RI Flg22 SIALGAADDAASNIRYGSSLRL SEQ ID NO: 442 Lysinibacillus sp.strain 13S34_air RI Flg22 SIAGLAADDSAGNIRLGSSLKG SEQ ID NO: 443Paenibacillus sp. strain HW567 RI Flg22 AIALGAADDSANNIRKGSSLRDSEQ ID NO: 444 Bacillus anthracis RI Flg22 AIALGAADDAASNIRKGSSLRNSEQ ID NO: 445 Bacillus anthracis RI Flg22 AIALGAADDAASNIRKGSSLRNSEQ ID NO: 446 Bacillus anthracis RI Flg22 AIALGAADDAASNIRKGSSLRNSEQ ID NO: 447 Bacillus anthracis RI Flg22 AIALGAADDAASNIRKGSSLRNSEQ ID NO: 448 Bacillus anthracis strain H9401 RI Flg22SIALGAADDSASNIRQGSSLKE SEQ ID NO: 449 Bacillus  megateriumstrain WSH-002 RI Flg22 SIALGAADDSARNIRYGSSLRE SEQ ID NO: 450Aneurinibacillus sp. XH2 Peptide Flg15 RI Flg15-8t4Q7 AIALGAADDKASNIRSEQ ID NO: 767 Modified FLG15- Bt4Q7; Syn01 Bacillus  thuringiensisstrain 4Q7 Peptide FlgII-28 RI FlgII-28-Bt.4Q7SAQNGKTNTGNASQNALDRMRLLINSVS SEQ ID NO: 451 Bacillus  thuringiensisstrain 4Q7 RI FlgII-28 SAQNGKTNTGNASQNALDRMRLLINSVS SEQ ID NO: 452Bacillus  thuringiensis, strain HD1002 RI FlgII-28SAQNGKTNTGNASQNALDRMRLLINSVS SEQ ID NO: 453 Bacillus  thuringiensis,strain HD-789 RI FlgII-28 SAQNGKTNTGNASQNALDRMRLLINSVS SEQ ID NO: 454Bacillus cereus strain G9842 RI FlgII-28 SHRNKNSNTGNASQVALDRMRQLINTVTSEQ ID NO: 455 Bacillus  thuringiensis serovar indiana strain HD521RI FlgII-28 ASKNENTNTGNASQNAIDRMRLLINSVS SEQ ID NO: 456 Bacillus thuringiensis strain CTC RI FlgII-28 AKQNDDTNTGNASQNALDRMRLLINSVSSEQ ID NO: 457 Bacillus  thuringiensis serovaryunnanensisstrain IEBC-T20001 RI FlgII-28 AAKNEDTNTGNASQNALDRMRLLINSVSSEQ ID NO: 458 Bacillus  thuringiensis serovar tolworthi RI FlgII-28LAVQNKDTNTGNASQNAIDRMRLLINSVS SEQ ID NO: 459 Bacillus cereus strain FM1RI FlgII-28 SDRNKNSNTGNASQVAVDRMRQLINTVT SEQ ID NO: 460 Bacillus cereusstrain FM1 RI FlgII-28 AQQNDATNTGNASQNAIDRMRLLINSVS SEQ ID NO: 461Bacillus  thuringiensis strain MC28 RI FlgII-28AAQNKDTNTGSASQNALDRMRLLINSVS SEQ ID NO: 462 Bacillus  bombysepticusstrain Wang RI FlgII-28 AAQNKDTNTGSASQNALDRMRLLINSVS SEQ ID NO: 463Bacillus  thuringiensis serovar kenyae RI FlgII-28AAQNKDTNTGSASQNALDRMRLLINSVS SEQ ID NO: 464 Bacillus  thuringiensisserovar kenyae RI FlgII-28 AKQNDGTNTGNASQNALDRMRLLINSVS SEQ ID NO: 465Bacillus cereus RI FlgII-28 NLSDRNKNSNTGNASQVALDRMRQLINT SEQ ID NO: 466Bacillus cereus RI FlgII-28 NLSDRNKNLNTGNASQVAVDRMRQLVNT SEQ ID NO: 467Bacillus  thuringiensis serovar finitimus strain YBT-020 RI FlgII-28ASKNSDTNTGNASQNAIDRMRLLINSVS SEQ ID NO: 468 Bacillus  thuringiensisserovar finitimus strain YBT-020 RI FlgII-28AAKNEATNTGNASQNALDRMRLLINSVS SEQ ID NO: 469 Bacillus cereus stain B4264RI FlgII-28 AKQNDSTNTGNASQNAIDRMRLLINSVS SEQ ID NO: 470 Bacillus thuringiensis serovar  nigeriensis RI FlgII-28AQQNDATNTGNASQNAIDRMRLLINSVS SEQ ID NO: 471 Bacillus  thuringiensisRI FlgII-28 SDRNKNSNTGNASQVALDRMRQLINMVT SEQ ID NO: 472 Bacillus thuringiensis serovar konkukian strain 97-27 RI FlgII-28AQQNDATNTGNASQNAIDRMRLLINSVS SEQ ID NO: 473 Bacillus  thuringiensisserovar konkukian strain 97-27 RI FlgII-28 SDRNTNSNTGNASQIAFDRMHQLINTVTSEQ ID NO: 474 Bacillus  thuringiensis serovar  thuringiensisstrain IS5056 RI FlgII-28 ASQNKDTNTGNASQNSIDRMRLLINSVS SEQ ID NO: 475Bacillus  thuringiensis serovar  thuringiensis strain IS5056 RI FlgII-28ASQNKDTNTGNASQNSIDRMRLLINSVS SEQ ID NO: 476 Bacillus  thuringiensisstrain Bt407 RI FlgII-28 ASQNKDTNTGNASQNSISRMRLLINSVS SEQ ID NO: 477Bacillus  thuringiensis serovar chinensis CT-43 RI FlgII-28AAQNENTNTGNASQNALDRMRLLINSVS SEQ ID NO: 478 Bacillus  thuringiensisserovar canadensis RI FlgII-28 AQQNEDTNTGNASQNSLDRMRLLINSVSSEQ ID NO: 479 Bacillus  thuringiensis serovar galleriae RI FlgII-28AQQNEDTNTGNASQNSLDRMRLLINSVS SEQ ID NO: 480 Bacillus weihenstephanensisRI FlgII-28 AKQNDGTNTGNASQNAIDRMRLLINSVS SEQ ID NO: 481 Bacillus thuringiensis serovar ostriniae RI FlgII-28 SDRNKNSNTDNSSQVALDRMRQLINAVTSEQ ID NO: 482 Bacillus  thuringiensis RI FlgII-28AKQNDDTNTGNASQNALDRMRLLINSVS SEQ ID NO: 483 Bacillus  thuringiensisRI FlgII-28 AKQNDDTNTGNASQNALDRMRLLINSVS SEQ ID NO: 484 Bacillus thuringiensis serovar pondicheriensis RI FlgII-28SDRNTNSNTGNASQIAFDRMHQLINTVT SEQ ID NO: 485 Bacillus  thuringiensisserovar Berliner RI FlgII-28 ASQNKDTNTGNASQNSIDRMRLLINSVS SEQ ID NO: 486Bacillus  thuringiensis serovar Berliner RI FlgII-28SDRNKSSNTGNASQVAVDRMRQLVNTVT SEQ ID NO: 487 Bacillus cereus strain Q1RI FlgII-28 AVQKDTNTGNASQNAIDRMRLLINSVS SEQ ID NO: 488 Bacillus cereusstrain Q1 RI FlgII-28 SDRNKNSNTSNASQIALDRMRQLINMVT SEQ ID NO: 489Bacillus  thuringiensis serovar morrisoni RI FlgII-28AKQNDSTNIGNASQNAIDRMRLLINSVS SEQ ID NO: 490 Bacillus  thuringiensisserovar  neoleonensis RI FlgII-28 AKQNDGTNTFNASQNAIDRMRLLINSVSSEQ ID NO: 491 Bacillus  thuringiensis serovar morrisoni RI FlgII-28AKQNDGTNTFNASQNAIDRMRLLINSVS SEQ ID NO: 492 Bacillus  thuringiensisserovar morrisoni RI FlgII-28 AAQNGNTNTFNASQNAIDRMRLLINSVSSEQ ID NO: 493 Bacillus  thuringiensis serovar jegathesan RI FlgII-28AAQNKDTNTGNASQNAIDRMRLLINSVS SEQ ID NO: 494 Bacillus cereusstain ATCC 10987 RI FlgII-28 from AAQNENTNTGNASQNALDRMRLLINSVSFlagellin A SEQ ID NO: 495 Bacillus  thuringiensis serovar monterreyRI FlgII-28 SDRNKNSNTDNASQVALDRMRQLVNTVT SEQ ID NO: 496 Bacillus cereusstrain NC7401 RI FlgII-28 AAKNENTNTGNASQNALDRMRLLINSVS SEQ ID NO: 497Bacillus cereus strain NC7401 RI FlgII-28 AAQNDSTNTGNASQNALDRMRLLINSVSSEQ ID NO: 498 Bacillus cereus strain AH820 RI FlgII-28AAKNENTNTGNASQNALDRMRLLINSVS SEQ ID NO: 499 Bacillus cereus AH187RI FlgII-28 AAKNENTNTGNASQNALDRMRLLINSVS SEQ ID NO: 500 Bacillus cereusRI FlgII-28 AKQNDGTNTGNASQNAIDRMRLLINSVS SEQ ID NO: 501 Bacillus cereusRI FlgII-28 AAQNKSTNTESASQNALDRMRLLINSVS SEQ ID NO: 502 Bacillus thuringiensis Strain HD-771 [51] RI FlgII-28AAQNKSTNTESASQNALDRMRLLINSVS SEQ ID NO: 503 Bacillus  thuringiensisserovar sotto [52] RI FlgII-28 AKQNDGTNTGNASQNAIDRMRLLINSVSSEQ ID NO: 504 Bacillus  thuringiensis serovar  Novosibirsk RI FlgII-28AAQNESTNTGNAQNALDRMRLLINSVS SEQ ID NO: 505 Bacillus  thuringiensisserovar londrina RI FlgII-28 SNRNKNSNTVNASQVALDRMRQLINTVT SEQ ID NO: 506Bacillus cereus strain E33L RI FlgII-28 AGQNKDTNTNASQNALDRMRLLINSVSSEQ ID NO: 507 Bacillus cereus strain E33L RI FlgII-28AAQNKDTNTGNASQNALDRMRLLINSVS SEQ ID NO: 508 Bacillus cereusstrain FRI-35 RI FlgII-28 SDRNKNSNTGNASQVALDRMRQLVNTVT SEQ ID NO: 509Bacillus cereus strain FRI-35 RI FlgII-28 AEIDTDKNTGNATQNAIDRMRLLINSVSSEQ ID NO: 510 Bacillus  thuringiensis RI FlgII-28AEIDTDKNTGNATQNAIDRMRLLINSVS SEQ ID NO: 511 Bacillus cereusstrain ATCC 4342 RI FlgII-28 SDRNKNSNTSNASQIALDRMRQLINMT SEQ ID NO: 512Bacillus  thuringiensis RI FlgII-28 AKQNDGTNTGNASQNAIDRMRLLINSVSSEQ ID NO: 513 Bacillus  thuringiensis RI FlgII-28ADLDETKNTGNGAQVVLERMRQLIDHTE SEQ ID NO: 514 Bacillus  aryabhattaiRI FlgII-28 SERDNGQNTGTAQTALDRMRLLINSVS SEQ ID NO: 515 Bacillus manliponensis RI FlgII-28 KARDDDTNTDTAAQVALERMRQVIAHTE SEQ ID NO: 516Lysinibacillus sp. strain BF-4 RI FlgII-28 KARDDDTNTDTAAQVALERMRQVIAHTESEQ ID NO: 517 Lysinibacillus sp. strain 13S34_air RI FlgII-28NLRDSGSYTGNSAQVALENMRQLMSHIE SEQ ID NO: 518 Paenibacillus sp.strain HW567 RI FlgII-28 ADQNEKTNTGNASQNALDRMRLLINSVS SEQ ID NO: 519Bacillus anthracis RI FlgII-28 ASQNEATNTGNSSQVAIDRMRTLINSVSSEQ ID NO: 520 Bacillus anthracis RI FlgII-28ASQNEATNTGNSSQVAIDRMRTLINSVS SEQ ID NO: 521 Bacillus anthracisRI FlgII-28 ASQNEATNTGNSSQVAIDRMRTLINSVS SEQ ID NO: 522Bacillus anthracis RI FlgII-28 ASQNEATNTGNSSQVIADRMRTLINSVSSEQ ID NO: 523 Bacillus anthracis strain H9401 RI FlgII-28ADLDETKNTGNGAQVVLERMRQLIDHTE SEQ ID NO: 524 Bacillus  megateriumstrain WSH-002 RI FlgII-28 AKKDKSYTGNAAQVALERMRQLMEHIE SEQ ID NO: 525Aneurinibacillus sp. XH2

Flg Sequences from Various Organisms

TABLE 5  Flagellin-associated Flg22 and Flg15  polypeptides from other organisms SEQ ID NO: Peptide-Amino AcidFlagellin (Flg22) ERLSSGLRINSAKDDAAGQAIA SEQ ID NO: 526 Escherichia coliFlagellin  AIAQGAADDKASNIRLGSSLRE (Retro-Inverso Flg22) SEQ ID NO: 527Escherichia coli Flagellin (Flg15) RINSAKDDAAGQAIA SEQ ID NO: 528Escherichia coli Flagellin  AIAQGAADDKASNIR (Retro-Inverso Flg15)SEQ ID NO: 529 Escherichia coli Flagellin (Flg22) QRLSTGSRINSAKDDAAGLQIASEQ ID NO: 530 Pseudomonas aeruginosa Flagellin  AIQLGAADDKASNIRSGTSLRQ(Retro Inverso Flg22) SEQ ID NO: 531 Pseudomonas aeruginosaFlagellin (Flg22) QRLSSGLRINSAKDDAAGLAIS SEQ ID NO: 532 Xanthomonas spp.X. compestris & X. citri Flagellin  SIALGAADDKASNIRLGSSLRQ(Retro Inverso Flg22) SEQ ID NO: 533 Xanthomonas spp.X. compestris & X. citri Flagellin (Flg22) QRLSSGLRINSAKDDAAGQAISSEQ ID NO: 534 Erwinia amylovora Flagellin  SIAQGAADDKASNIRLGSSLRQ(Retro Inverso Flg22) SEQ ID NO: 535 Erwinia amylovora Flagellin (Flg22)TRLSSGKRINSAADDAAGLAIS SEQ ID NO: 536 Burkholderia phytofirmansFlagellin  SIALGAADDAASNIRKGSSLRT (Retro Inverso Flg22) SEQ ID NO: 537Burkholderia phytofirmans Flagellin (Flg22) NRLSSGKRINTAADDAAGLAISSEQ ID NO: 538 Burkholderia ubonensis Flagellin  SIALGAADDAATNIRKGSSLRN(Retro Inverso Flg22) SEQ ID NO: 539 Burkholderia ubonensisFlagellin (Flg22) TRLSSGLKINSAKDDAAGLQIA SEQ ID NO: 540Pseudomonas syringae Flagellin  AIQLGAADDKASNIKLGSSLRT(Retro Inverso Flg22) SEQ ID NO: 541 Pseudomonas syringaeFlagellin (Flg11-28) ESTNILQRMRELAVQSRNDSNS (SEQ ID NO: 751) ATDREAPseudomonas syringae Flagellin  AERDTASNSDNRSQVALERMRQ(Retro Inverso Flg11-28) LINTSE (SEQ ID NO: 768) Pseudomonas syringae

Sequences that Assist in Directing Flagellins or Flagellin-AssociatedPolypeptides to the Plant

The signature, signal anchor sorting and secretion sequences can be usedseparately or together in combination with any of the flagellin orflagellin-associated polypeptides as described herein. These assistancesequences are useful for the efficient delivery of the flagellinpolypeptides to the plant cell membrane surface. Other assistancesequences can also assist with the translocation of the Flg polypeptidefragment across the plasma membrane. Delivery of flagellins andflagellin-associated polypeptides to the plasma membrane surface of aplant (or plant part) can contribute to downstream signalling processesand result in beneficial outcomes to a plant or a plant part, such asenhanced plant health and productivity.

The polypeptide can further comprise an assistance polypeptide.

The assistance polypeptide can comprise a signature polypeptide, and anamino acid sequence of the signature polypeptide can comprise any one ofSEQ ID NOs: 542-548, listed in Table 6, or any combination thereof. Forexample, the amino acid sequence of the signature polypeptide cancomprise SEQ ID NO: 542.

The assistance polypeptide can comprise a signal anchor sortingpolypeptide, and an amino acid sequence of the signal anchor sortingpolypeptide can comprise any one of SEQ ID NOs: 549-562, listed in Table7, or any combination thereof. For example, the amino acid sequence ofthe signal anchor sorting polypeptide can comprise SEQ ID NO: 549.

The flagellin or flagellin-associated polypeptide can be producedrecombinantly by a microorganism. For example, the microorganism cancomprise a Bacillus, a Pseudomonas, a Paenibacillus, Aneurinibacillus ora Lysinibacillus.

The assistance polypeptide can comprise a secretion polypeptide, and anamino acid sequence of the secretion polypeptide can comprise any one ofSEQ ID NOs: 563-570, or any combination thereof. For example, the aminoacid sequence of the secretion polypeptide can comprise SEQ ID NO: 563.

These three types of assistance sequences are further described in Table6 (N-terminal signature sequences), Table 7 (signal anchor sortingsequences) and Table 8 (secretion sequences).

Also provided are “assistance” sequences having conserved signature(Table 6; SEQ ID NOs: 542-548), signal anchor sorting (Table 7; SEQ IDNOs: 549-562) and secretion (Table 8; SEQ ID NOs: 563-570) sequences incombination with any of the flagellin-associated polypeptides asdescribed herein. Particularly useful are combinations of the signature,signal anchor sorting and secretion assistance sequences with the nativeL-Flg polypeptides (Table 3. SEQ ID NOs: 226-375) or any of the retroinverso Flg22 polypeptides (Table 4. SEQ ID NOs: 376-525) for providingefficient delivery of the Flg polypeptides to the extracellular plantmembrane surface, such as the surface of a plant or plant part.

N-Terminal Signature Sequences

Amino acid “signature” sequences conserved within Bacillus,Lysinibacillus, Paenibacillus or Aneurinibacillus bacteria (genera) andother Eubacterial generas can function in targeting flagellinpolypeptides to the appropriate Flg-associated receptor protein(s), suchas FLS receptors that have an exposed binding site at the plant cellmembrane surface and can be used to enhance Flg polypeptide-receptorbinding leading to an increased activation potential of theFlg-associated receptor(s). Flagellin signature sequences as identifiedin Table 6 are useful for targeting and stably delivering the Flgpolypeptides for binding to the FLS or FLS-like receptor(s) thereforeincreasing the contact and binding between the membrane receptor and theFlg polypeptide.

Conserved N-terminal signature sequences (SEQ ID NO: 542-548) can beused in combination with any of the flagellin-associated polypeptides asdescribed herein. Of particular utility are the signature sequences usedin combination with the native L-Flg polypeptides (L-Flg22 SEQ ID NOs:226-300; L-FlgII-28 SEQ ID NOs: 301-375) or any of the retro inversoD-Flg polypeptides (D-Flg22 SEQ ID NOs: 376-450; FlgII-28 SEQ ID NO:451-525) or any of the other Flg-associated sequences provided in Table5 (SEQ ID NOs: 526-541) to provide efficient delivery of theFlg-associated polypeptides to the plant membrane surface.

Signature sequences assist with Flg22 and FlgII-28 bioactive primingpolypeptide sequences in binding to the appropriate Flg-associatedreceptor(s) in order to activate the receptor(s) making it functionallyactive.

TABLE 6 Flagellin-associated N-terminal  signature sequencesFlagellin Signature SEQ ID NO: Sequences SEQ ID NO: 542 GFLNSEQ ID NO: 543 WGFLI SEQ ID NO: 544 MGVLN SEQ ID NO: 545 GVLNSEQ ID NO: 546 WGFFY SEQ ID NO: 547 LVPFAVWLA SEQ ID NO: 548 AVWLA

N-Terminal Signal Anchor Sorting Sequences

Amino acid “signal anchor sorting” sequences conserved within Bacillus,Lysinibacillus, Aneurinibacillus and Paenibacillus genera and otherEubacterial generas' bacteria can function in anchoring and localizingthe flagellin-associate polypeptides to the plant cell membrane surfaceand assist in high affinity binding to the appropriate Flg-associatedreceptor(s) thereby increasing the activation potential of the boundreceptor(s).

Conserved signal anchor sequences (SEQ ID NO: 549-562; Table 7) arelocated downstream of the pre-cleaved or full-length coding or partialcoding flagellin sequences, for example, as described herein (SEQ IDNOs: 1-75; Table 1).

The signal anchor sorting domains as described herein are useful inmembrane attachment. They can be used to aid in the localization andbinding of Flg-associated polypeptides to a surface membrane receptorand have some functional similarity at the amino acid level to proteinsthat are endosomal (vesicular) trafficked or destined for targeting tothe secretory pathway. Such signal anchor sorting sequences as describedherein that are useful for anchoring the Flg bioactive primingpolypeptides to the plant cell membrane are also used to enhance themembrane integration of the bioactive priming Flg polypeptides into theplant cell.

Such sequences as described in Table 7 may further be functionallyannotated as import receptor signal anchor sequences, which can be usedto improve targeting or delivery and efficient membrane anchoring ofFlg-associated polypeptides to a plant and assist with membraneintegration into the cytosol of the plant cell.

Combining the signal anchor sequences (SEQ ID NOs: 549-562; Table 7)with any of the flagellins or flagellin-associated bioactive primingpolypeptides as described herein is useful to facilitate the attachmentand import of these flagellin-associated polypeptide(s) into the plant.

Such signal anchor sorting sequences can be used in combination with theFlg-associated polypeptides, and are useful for targeting, efficientmembrane anchoring, membrane integration and Golgi-to-lysosomal/vacuolartrafficking. The signal anchor sorting sequences are used to stablydeliver the Flg polypeptides to the plant membrane surface andintegrally incorporate them into the plant.

Such sequences as described herein contain di-leucine amino acids thatare referenced to confer endocytosis functionalities in plant systems(Pond et al. 1995, “A role for acidic residues in di-leucine motif-basedtargeting to the endocytic pathway”, Journal of Biological Chemistry270: 19989-19997, 1995).

Such signal anchor sorting sequences as described can also be used toefficiently deliver systemic signals to infection sites and stimulate aplant's innate immunity in plant cells.

TABLE 7  Flagellin-associated signal  anchor sorting sequencesSEQ ID NO: Signal Anchor Sequence SEQ ID NO: 549 LLGTADKKIKIQSEQ ID NO: 550 LLKSTQEIKIQ SEQ ID NO: 551 LLNEDSEVKIQ SEQ ID NO: 552LGVAANNTQ SEQ ID NO: 553 LLRMRDLANQ SEQ ID NO: 554 LQRMRDVAVQSEQ ID NO: 555 LLRMRDISNQ SEQ ID NO: 556 LLRMRDIANQ SEQ ID NO: 557LQKQIDYIAGNTQ SEQ ID NO: 558 LLIRLPLD SEQ ID NO: 559 QRMRELAVQSEQ ID NO: 560 TRMRDIAVQ SEQ ID NO: 561 TRMRDIAVQ SEQ ID NO: 562QRMRELVVQ

C-Terminal Secretion Sequences

Conserved sequences located in the C-terminus of flagellin(s) arefurther described as secretion sequences (SEQ ID NO: 563-570; Table 8).

Conserved sequences were identified in the C-terminus of the Bacillus,Lysinibacillus, and Paenibacillus bacteria (genera) and otherEubacterial genera derived flagellin proteins and comprise 6 aminoacids, for example LGATLN, LGAMIN, or LGAMIN. These sequences werefunctionally annotated using BLAST against the bacterial databases asmotifs that have highest homology to secretion polypeptides. The 6 aminoacid conserved polypeptides identified were found most similar to thosefound in type III secretion systems in E. coli. Type III export systemshave been cited to be involved in the translocation of polypeptidesacross the plant cell membrane. The filament assembly of flagellin isdependent on the availability of flagellins to be secreted and mayrequire chaperones that assist in the secretory process.

These secretion polypeptides as described herein may be used incombination with any of the flagellin-associated polypeptides asdescribed herein to deliver these polypeptides/peptides into the cytosolof the host plant thus providing beneficial outcomes to a plant.

TABLE 8  C-terminal flagellin-associated   secretion sequencesFlagellin Secretion SEQ ID NO: polypeptides SEQ ID NO: 563 LGATLNSEQ ID NO: 564 LGATQN SEQ ID NO: 565 LAQANQ SEQ ID NO: 566 LGAMINSEQ ID NO: 567 LGSMIN SEQ ID NO: 568 MGAYQN SEQ ID NO: 569 LGAYQNSEQ ID NO: 570 YGSQLN

The signature (SEQ ID NO: 542-548; Table 6), signal anchor sorting (SEQID NO: 549-562; Table 7) and secretion (SEQ ID NO: 563-570; Table 8)sequences as provided herein can be used with any of the flagellinpolypeptides or the flagellin-associated polypeptides to promote growthand provide health and protective benefits to a plant or a plant part.

Modification of Flg Polypeptide Sequences Function

Any of the L or D Flg-associated sequences provided in Tables 3, 4 or 5can be similarly modified as fused to any of the assistance sequences asdescribed in Table 6-8. For one example, fusion of any of theseassistance sequences will present a modification to the Bt.4Q7Flg22bioactive priming polypeptide sequence identified as SEQ ID NO: 226.

Mutations to Flg-Associated Polypeptides to Increase Responsiveness toReactive Oxygen Species or Polypeptide Stability

The polypeptide can comprise a mutant flagellin or flagellin-associatedpolypeptide.

The mutant flagellin or flagellin-associated polypeptide can be derivedfrom a Bacillus, a Lysinibacillus, a Paenibacillus, or anAneurinibacillus genus bacterium. Other polypeptides from otherEubacterial classes, including Enterobacteraciae, can also be used inthe same fashion. Other generas of interest include Pseudomonas,Escherichia, Xanthomonas, Burkholderia, Erwinia, and others.

The amino acid sequence of the flagellin or flagellin-associatedpolypeptide can comprise any one of SEQ ID NOs: 226, 289, 290, 291, 293,294, 295, 300, 437, 532, 534, 536, 538, 540, 571-586 and 751-768. Forexample, the amino acid sequence of the flagellin orflagellin-associated polypeptide can comprise any one of SEQ ID NOs:226, 293, 295, 300, 540, 571, 574 and 752, or any combination thereof.

Any bioactive priming polypeptide, whether naturally occurring ornon-natural, can be further modified via chemical modification toincrease performance as well as stability of the polypeptides. Suchbioactive priming polypeptides include flagellin polypeptides, retroinverso polypeptides, harpin derived polypeptides, harpin-like derivedpolypeptides, EF-Tu polypeptides, thionin polypeptides, RHPPpolypeptides, and PSK polypeptides. Specific sequences that can bechemically modified include SEQ ID NOs: 226-592, 594-601, 603-749, and751-766.

These bioactive priming polypeptides can also be conjugated to othermoieties, including a plant binding domain and a polypeptide, a plantpart binding domain and a polypeptide, and other carriers such as oils,plastics, beads, ceramic, soil, fertilizers, pellets, and moststructural materials.

The flagellin or flagellin-associated polypeptide can be modifiedchemically on its N or C terminus. Common modification of the N andC-termini include: acetylation, lipid addition, urea addition,pyroglutamyl addition, carbamate addition, sulfonamide addition,alkylamide addition, biotinylation, phosphorylation, glycosylation,PEGylation, methylation, biotinylation, acid addition, amide addition,ester addition, aldehyde addition, hydrazide addition, hydroxyamic acidaddition, chloromethyl ketone addition, or addition of purificationtags. These tags can increase activity of the polypeptides, increasestability, add protease inhibitor abilities to the polypeptides, blockproteases directly, allow for tracking, and help in binding to planttissues.

The flagellin or flagellin-associated polypeptide can be modified viacrosslinking or cyclization. Crosslinking can bind polypeptides eitherto each other or to a secondary surface or moiety to help in delivery orstability of the polypeptides. Cyclization can be performed, forexample, to both increase activity of the polypeptide as well as preventprotease interaction with the polypeptide.

Sequence modifications or mutations can be made to any amino acidsequence(s) as described in Tables 4 and 5 and replaced with any of the20 standard amino acid sequences known in nature or replaced with anonstandard or non-canonical amino acid sequence, such asselenocysteine, pyrrolysine, N-formylmethione, etc. For example,modifications or mutations can be made to the internal sequences asshown in SEQ ID NO: 571, to the C-terminis as shown in SEQ ID NO: 572 orSEQ ID NO: 753, or to the N terminus as shown in SEQ ID NO: 573 toproduce Flg polypeptides with enhanced ROS activates and increasedfunctionality in a plant or plant part. Modified polypeptides also canbe truncated at the N or C terminus as shown in SEQ ID NO: 752(N-terminus truncation) to further increase functionality in a plant orplant part. Table 9A summarizes flagellin polypeptides identified thatprovide modified ROS activity.

TABLE 9A Flagellin polypeptides Flg22 identified from Bacillus or other bacteriawith mutations that provide modified ROS activity SEQ ID NO:Peptide Flg22 Flg22-Bt4Q7

SEQ ID NO:-571 Bacillus thuringiensis strain 4Q7Modified FLG22-Bt4Q7 (S13K); Syn01 Flg22-Bt4Q7 DRLSSGKRINSASDDAAGLQIASEQ ID NO: 572 Bacillus thuringiensis strain 4Q7Modified FLG22-Bt4Q7 (A20Q); Syn02 Flg22-Bt4Q7

SEQ ID NO: 573 Bacillus thuringiensis strain 4Q7Modified FLG22-Bt4Q7 (D1Q); Syn03 Flg22-Bt4Q7

SEQ ID NO: 574 Bacillus thuringiensis strain 4Q7Modified FLG22-Bt4Q7 (D1N); Syn06 Caballeronia megalochromosomata

SEQ ID NO: 575 Flg22-Bt4Q7 DRLSSGYRINSASDDAAGLAIA SEQ ID NO: 576Bacillus thuringiensis strain 4Q7 Modified FLG22-Bt4Q7 (K7Y); Syn07Flg22-Bt4Q7 DRLSSGFRINSASDDAAGLAIA SEQ ID NO: 577 Bacillus thuringiensisstrain 4Q7 Modified FLG22-Bt4Q7 (K7F); Syn08 Flg22-Br4Q7

SEQ ID NO: 578 Bacillus thuringiensis Modified FLG22-Bt4Q7 (A16P); Syn05Flg22-Bt4Q7

SEQ ID NO: 579 Bacillus thuringiensis strain 4Q7Modified FLG22-Bt4Q7 (K7Q); Syn09 Flg22-Br4Q7

SEQ ID NO: 753 Bacillus thuringiensis strain 4Q7Modified FLG22-Bt4Q7 (D15P); Syn04 Flg15-Br4Q7

SEQ ID NO: 752 Bacillus thuringiensis N-term Truncated Syn01 Bm.Flg22-B1NRLSSGKQINSASDDAAGLAIA Bacillus manliponensis SEQ ID NO: 290 Ba.Flg22-B2NRLSSGKRINSAADDAAGLAIA Bacillus anthracis SEQ ID NO: 295 Bc.Flg22-B3DRLSSGKRINNASDDAAGLAIA Bacillus cereus SEQ ID NO: 294 A. spp.Flg22-B4ERLSSGYRINRASDDAAGLAIS Aneurinibacillus spp. XH2 SEQ ID NO: 300Ba.Flg22-B5 EKLSSGQRINSASDDAAGLAIS Bacillus aryabhattai SEQ ID NO: 289P spp.Flg22-B6 GKLSSGLRINGASDDAAGLAIS Paenibacillus spp. strain HW567SEQ ID NO: 293 L spp.Flg22-L1 LRLSSGYRINSAADDAAGLAIS Lysinibacillus spp.SEQ ID NO: 291 L spp.Flg22-L2 EKLSSGLRINRAGDDAAGLAIS Lysinibacillus spp.SEQ ID NO: 580 L spp.Flg22-L3 EKLSSGYKINRASDDAAGLAIS Lysinibacillus spp.SEQ ID NO:-581 L spp.Flg22-L4 LRISSGYRINSAADDPAGLAISLysinibacillus spp. SG9 SEQ ID NO: 582 Lf.Flg22-L5LRISTGYRINSAADDPAGLAIS Lysinibacillus fusiformis SEQ ID NO: 583Lm.Flg22-L6 EKLSSGFRINRAGDDAAGLAIS Lysinibacillus macroidesSEQ ID NO: 584 Lx.Flg22-L6 EKLSSGYKINRAGDDAAGLAIS Lysinibacillusxylanilyticus SEQ ID NO: 585 Pa.Flg22 QRLSTGSRINSAKDDAAGLQIAPseudomonas aeruginosa SEQ ID NO: 530 Ec.Flg22 ERLSSGLRINSAKDDAAGQAIAEscherichia coli SEQ ID NO: 586 Xcc.Flg22 QRLSSGLRINSAKDDAAGLAISXanthomonas campestris pv campestris strain 305 or(Xanthomonas citri pv. citri) SEQ ID NO: 532 Ea.Flg22 QRLSSGLRINSAKDDAAGQAIS Erwinia amylovora SEQ ID NO: 534 Bp.Flg22TRLSSGKRINSAADDAAGLAIS Burkholderia phytofirmans strain PsJNSEQ ID NO: 536 Bu.Flg22 NRLSSGKRINTAADDAAGLAIS Burkholderia ubonensisSEQ ID NO: 538 Ps.Flg22 TRLSSGLKINSAKDDAAGLQIAPseudomonas syringae pv. actinidiae ICMP 19096 SEQ ID NO: 540

Core Active Domain of Flg22

The underlined portions of the sequences in Table 9A represent the coreactive domain of Flg22. This core domain comprises, for example, SEQ IDNO: 754 with up to one, two or three amino acid substitutions(represented by SEQ ID NOs 755-765) that can promote growth, diseasereduction and/or prevention in crops and ornamental plants. For ease ofreference, this core domain is represented as the consensus sequencehaving the SEQ ID NO: 766. The various native and mutant Flg22polypeptides comprising SEQ ID NOs 754-765 are described along with theconsensus sequence in Table 9B, below. Therefore, the polypeptides canfurther comprise a core sequence. The core sequence can comprise any oneof SEQ ID NOs 754-766.

The polypeptide can also comprise any polypeptide comprising any one ofSEQ ID NOs 1-753 or 767 to 768 wherein the polypeptide further comprisesthe core sequence comprising any one of SEQ ID NOs: 754-766. Theinclusion of the core sequence in the polypeptide or full-length proteinof dissimilar function can increase the bioactive priming activity ofthe polypeptide.

Table 9B Flg22 core sequence with variants. FLG22 corePolypeptides comprising SEQ ID NO:  sequence core sequenceSEQ ID NO: 754 RINSASDD SEQ ID NO: 226-229 SEQ ID NO: 289 SEQ ID NO: 299SEQ ID NO: 536 SEQ ID NO: 572-579 SEQ ID NO: 755 RINNASDDSEQ ID NO: 231-234 SEQ ID NO: 236-240 SEQ ID NO: 243-246 SEQ ID NO: 248SEQ ID NO: 250-256 SEQ ID NO: 258-259 SEQ ID NO: 261 SEQ ID NO: 263SEQ ID NO: 265-270 SEQ ID NO: 272-280 SEQ ID NO: 282-283SEQ ID NO: 285-286 SEQ ID NO: 288 SEQ ID NO: 294 SEQ ID NO: 756 QINSASDDSEQ ID NO: 290 SEQ ID NO: 757 RINSAADD SEQ ID NO: 291-292SEQ ID NO: 295-298 SEQ ID NO: 582-583 SEQ ID NO: 536 SEQ ID NO: 582-583SEQ ID NO: 758 RINGASDD SEQ ID NO: 293 SEQ ID NO: 759 RINRASDDSEQ ID NO: 300 SEQ ID NO: 760 RINSAKDD SEQ ID NO: 526 SEQ ID NO: 528SEQ ID NO: 530 SEQ ID NO: 532 SEQ ID NO: 534 SEQ ID NO: 571SEQ ID NO: 586 SEQ ID NO: 761 RINTAADD SEQ ID NO: 538 SEQ ID NO: 762KINSAKDD SEQ ID NO: 540 SEQ ID NO: 763 RINRAGDD SEQ ID NO: 580SEQ ID NO: 584 SEQ ID NO: 764 KINRASDD SEQ ID NO: 581 SEQ ID NO: 765KINRAGDD SEQ ID NO: 585 SEQ ID NO: 766 (R/Q/K)IN Consensus of SEQ ID NO:(S/N/G/R/T) 755-765 (sequences A(S/A/K/G)DD identified in this table)

Harpin or Harpin-Like Polypeptides

The polypeptide can include a harpin or harpin-like polypeptide.

The amino acid sequence of the harpin or harpin-like polypeptide cancomprise SEQ ID NOs: 587-592 and 594-597 (Tables 10 and 11),

The harpin or harpin-like polypeptides can be derived from Xanthomonasspecies or diverse bacteria genera including Pantoea sesami, Erwiniagerudensis, Pantoea sesami, or Erwinia gerudensis

Additional Harpin-like bioactive priming polypeptides can be derivedfrom the full length HpaG-like protein from Xanthamonas citri comprisingSEQ ID NO: 593.

Application of HpaG-like polypeptides using the native L-harpin-likesequence (SEQ ID NO: 587) or retro inverso D-harpin-like sequence (SEQID NO: 588) bioactive priming polypeptides forms as represented inTables 10 or 11 are useful to increase growth and immune responses inplants when applied either exogenously or endogenously to a plant orplant part. The retro-inverso HpaG-like (e.g. SEQ ID NO: 588) bioactivepriming polypeptide is particularly useful to enhance the activity andstability of the HpaG-like polypeptide when applied to plants grownunder or exposed to conditions of abiotic stress. The retro-inversoHpaG-like form can be used to enhance growth and protection responses inplants grown under such environments.

TABLE 10  Harpin-like (HpaG-like) Peptide Sequence  SEQ ID NO:Amino Acid Harpin-like (HpaG-like) NQGISEKQLDQLLTQLIMALLQQSEQ. ID NO: 587 Xanthomonas species MW 2626.35 Da Harpin-like QQLLAMILQTLLQDLQKESIGQN (Retro-Inverso HpaG-like) SEQ. ID NO: 588Xanthomonas species MW 2626.35 Da Harpin-like (HpaG-like) LDQLLTQLIMALSEQ. ID NO: 589 Xanthomonas species MW 2626.35 Da Harpin-like LAMILQTLLQDL (Retro-Inverso HpaG-like) SEQ. ID NO: 590Xanthomonas species MW 2626.35 Da Harpin-like (HpaG-like)SEKQLDQLLTQLIMALLQQ SEQ. ID NO: 591 Xanthomonas species MW 2626.35 DaHarpin-like  QQLLAMILQTLLQDLQKES (Retro-Inverso HpaG-like)SEQ. ID NO: 592 Xanthomonas species MW 2626.35 Da HpaG-Like ProteinMMNSLNTQLGANSSFFQVDPSQN SEQ. ID NO: 593 TQSGSNQGNQGISEKQLDQLLTQXanthamonas citri LIMALLQQSNNAEQGQGQGQGGD SGGQGGNRQQAGQSNGSPSQYTQMLMNIVGDILQAQNGGGFGGGFG GGFGGGLGTSLGTSLGTSLASDT GSMQ

TABLE 11  HpaG-like Homologs from diverse bacterial genera SEQ ID NO:Peptide amino acid HpaG Homolog QLEQLMTQLRARLCRLMAM Active FractionSEQ ID NO: 594 Pantoea sesami HpaG Homolog QLEQLMTQLRARLKRLMAMActive Fraction SEQ. ID NO: 595 Erwina gerudensis Retro InversoMAMLRCLRARLQTMLQELQ HpaG Homolog Active Fraction SEQ. ID NO: 596Pantoea sesami Retro Inverso MAMLRKLRARLQTMLQELQ HpaG HomologActive Fraction SEQ. ID NO: 597 Erwinia gerudensis

Phytosulfokine (PSKα) Polypeptides

The polypeptide can comprise the PSK polypeptide.

The amino acid sequence of the PSK polypeptide can comprise SEQ ID NOs:598-599.

Phytosulfokine alpha (PSKα) was originally derived from Arabidopsisthaliana and is a sulfonated bioactive priming polypeptide. The PSKabioactive priming polypeptide(s) are in Table 11.

PSKα is provided either as a synthetic polypeptide or a naturalpolypeptide that is expressed in a recombinant microorganism, purifiedand used in agricultural formulations for applications to plants orplant parts.

TABLE 12 Phytosulfokine alpha (PSKα), sulfonated bioactive priming polypeptides provided as  natural and retro-inverso amino acid sequences SEQ ID NO:Peptide Sequence Amino Acid Phytosulfokine (PSKα)Tyr(SO₃H)-I-Tyr(SO₃H)-TQ SEQ. ID NO: 598 Arabidopsis thaliana MW 845 DaPhytosulfokine  QT-Tyr(SO₃H)-I-Tyr(SO₃H) (Retro Inverso PSKα)SEQ ID NO: 599 Arabidopsis thaliana MW 845 Da

Root Hair Promoting Polypeptide (RHPP)

The polypeptide can comprise a RHPP

The amino acid sequence of the RHPP can comprise SEQ ID NO: 600-601 and603-606. For example, the amino acid sequence of the RHPP can compriseSEQ ID NO: 600.

A combination of the polypeptide comprising an RHPP and a polypeptidecomprising a flagellin or flagellin associated polypeptide is alsoprovided. The flagellin or flagellin associated polypeptide can compriseany one of SEQ ID NO: 226, 752, and 571. In some instances, thepolypeptide comprises an RHPP comprising SEQ ID NO: 600 and a flagellincomprising SEQ ID NO: 226.

The polypeptide can comprise the PSK polypeptide, the RHPP, the harpinor harpin-like polypeptide, or a combination thereof.

Additional RHPP bioactive priming polypeptides can be derived from thefull length Kunitz Trypsin Inhibitor protein from Glycine max comprisingSEQ ID NO: 602. The RHPP polypeptide can be modified via C-terminalamidation, N-terminal acetylation or other modification. The RHPPbioactive priming polypeptide can be obtained through addition of crudeprotease digest of kunitz trypsin inhibitor and/or soybean meal.

RHPP originally derived for soybean (Glycine max) can be provided, forexample, as a foliar application to produce beneficial phenotypes incorn, soybean and other vegetables.

TABLE 13 Amino acid sequence for RHPP forward and  sequencesretro-inverso SEQ ID NO: Peptide Sequence Amino AcidRoot Hair Promoting  GGIRAAPTGNER Peptide (RHPP) SEQ. ID NO: 600Glycine max MW 1198.20 Da Root Hair Promoting  RENGTPAARIGG Peptide (Retro Inverso RHPP) SEQ. ID NO: 601 Glycine max MW 1198.20 DaKunitz Trypsin  MKSTIFFALFLFCAFTTSYLPSAIADFV InhibitorLDNEGNPLENGGTYYILSDITAFGGIRA SEQ. ID NO: 602APTGNERCPLTVVQSRNELDKGIETIIS Glycine Max SPYRIRFIAEGHPLSLKFDSFAVIMLCVGIPTEWSVVEDLPEGPAVKIGENKDAMD GWFRLERVSDDEFNNYKLVFCPQQAEDDKCGDIGISIDHDDGTRRLVVSKNKPLVV QFQKLDKESLAKKNHGLSRSE

TABLE 14  Homologs of RHPP from Glycine spp. Peptide Sequence SEQ ID NO: Amino Acid Homolog RHPP GGIRATPTENER SEQ ID NO: 603Glycine max Homolog RHPP GGIRVAATGKER SEQ ID NO: 604Glycine max/Glycine soja

The polypeptide can include a retro inverso (RI) RHPP.

The retro inverso RHPP can comprise SEQ ID NOs: 601, 605 or 606.

The retro inverso (RI) RHPP can be modified via C-terminal amidation orN-terminal acetylation.

TABLE 15 Retro inverso amino acid sequences for homologs of RHPP from Glycine spp. Peptide Sequence  SEQ ID NO:Amino Acid Homolog RHPP RENETPTARIGG SEQ. ID NO: 605 Glycine maxHomolog RHPP REKGTAAVRIGG SEQ. ID NO: 606 Glycine max/Glycine soja

Elongation Factor Tu (EF-Tu) Polypeptides

The polypeptide can comprise an EF-Tu polypeptide.

Peptides derived from elongation factor Tu (EF-Tu) can be usedseparately or in combination with the other bioactive primingpolypeptides as described herein such as in combination with Flg22polypeptides to provide multiple modes of defense against pathogenicorganisms, generally bacterial and fungal microorganisms but alsoincluding other infection agents, such as viruses.

Table 16 provides preferred N-terminal polypeptides derived from variousEF-Tu bioactive priming polypeptides selected from both plants andbacteria. The EF-Tu derived polypeptides can be any length from 18 to 26amino acids or less than 26 amino acids in length. Table 17 furtherprovides retro-inverse (all-D) versions of EF-Tu polypeptides derivedfrom bacteria and algae.

The amino acid sequence of the EF-Tu polypeptide can comprise and one ofSEQ ID NOs: 607-640.

The amino acid sequence of the EF-Tu polypeptide can comprise SEQ ID NO:616 or 617.

The EF-Tu polypeptide can be modified via N-terminal acetylation. Forexample, the EF-Tu polypeptide can be modified via N-terminalacetylation and comprise any of SEQ ID NOs: 607, 608, 610, 611, 613,614, 616, 617, 619, or 622.

TABLE 16 N-terminal acetylated and central polypeptides derived from elongation factors (EF-Tu) existingin plant, bacterial and algae species Length amino SEQ ID NO: acidsPeptide amino acid Chloroplastic EF-Tu 18 Ac-ARGKFERKKPHVNIGTIGSEQ ID NO: 607 (acetylated) Arabidopsis lyrata Chloroplastic EF-Tu 26Ac-ARGKFERKKPHVNIGTIG SEQ ID NO: 608 HVDHGKTT (acetylated)Arabidopsis lyrata Chloroplastic EF-Tu 50 EKPNVKRGENKWVDKIYELMDSVDSSEQ ID NO: 609 YIPIPTRQTELPFLLAVEDVFSITG Arabidopsis lyrataN-terminus of EF-Tu 18 Ac-ARQKFERTKPHINIGTIG SEQ ID NO: 610 (acetylated)Euglena gracilis N-terminus of EF-Tu 26 Ac-ARQKFERTKPHINIGTIGHVDHSEQ ID NO: 611 GKTT (acetylated) Euglena gracilis EF-Tu fragment 50KNPKITKGENKWVDKILNLMDQVDS SEQ ID NO: 612 YIPTPTRDTEKDFLMAIEDVLSITGEuglena gracilis N-terminus of EF-Tu 18 Ac-AKGKFERTKPHVNVGTIGSEQ ID NO: 613 (acetylated) Acidovorax avenae N-terminus of EF-Tu 26Ac-AKGKFERTKPHVNVGTIGHVDH SEQ ID NO: 614 GKTT (acetylated)Acidovorax avenae EF-Tu fragment 50 KLALEGDKGPLGEQAIDKLAEALDTSEQ ID NO: 615 YIPTPERAVDGAFLMPVEDVFSISG Acidovorax spp.N-terminus of EF-Tu 18 Ac-AKAKFERSKPHVNIGTIG SEQ ID NO: 616 (acetylated)Bacillus cereus N-terminus of EF-Tu 26 Ac-AKAKFERSKPHVNIGTIGHVDHSEQ ID NO: 617 GKTT (acetylated) Bacillus cereus EF-Tu fragment 50SALKALQGEAEWEEKIIELMAEVDA SEQ ID NO:618 YIPTPERETDKPFLMPIEDVFSITGBacillus cereus N-terminus of EF-Tu 26 Ac-AKGKFERTKPHVNVGTIGHVDHSEQ. ID NO: 619 GKTT (acetylated) Burkholderia spp. EF-Tu fragment 50KLALEGDTGELGEVAIMNLADALDT SEQ. ID NO: 620 YIPTPERAVDGAFLMPVEDVFSISGBurkholderia spp. EF-Tu fragment 50 RLALDGDQSEIGVPAILKLVDALDTSEQ ID NO: 621 FIPEPTRDVDRPFLMPVEDVFSISG Xanthomonas campestrisN-terminus of EF-Tu 26 Ac-AKEKFERSKPHVNVGTIGHVDH SEQ. ID NO: 622 GKTT(acetylated) Pseudomonas spp. EF-Tu 50 MALEGKDDNEMGTTAVKKLVETLDSSEQ. ID NO: 623 YIPEPERAIDKPFLMPIEDVFSISG Pseudomonas spp.

TABLE 17 Retro Inverso polypeptides derived from elongation factors (EF-Tu) existing in bacterial and algae species Length aminoSEQ ID NO: acids Peptide amino acid RI Chloroplastic  18GITGINVHPKKREFKGRA EF-Tu SEQ. ID NO: 624 Arabidopsis lyrataRI Chloroplastic  26 TTKGHDVHGITGINVHPKK EF-Tu REFKGRA SEQ. ID NO: 625Arabidopsis lyrata RI Chloroplastic  50 GTISFVDEVALLFPLETQR EF-TuTPIPIYSDVSDMLEYIKDV SEQ ID NO: 626 WKNEGRKVNPKE Arabidopsis lyrataRI N-terminus of  18 GITGINIHPKTREFKQRA EF-Tu SEQ. ID NO: 627Euglena gracilis RI N-terminus of  26 TTKGHDVHGITGINIHPKT EF-Tu REFKQRASEQ. ID NO: 628 Euglena gracilis RI EF-Tu fragment 50GTISLVDEIAMLFDKETDR SEQ ID NO: 629 TPTPIYSDVQDMLNLIKDV Euglena gracilisWKNEGKTIKPNK RI N-terminus of  18 GITGVNVHPKTREFKGKA EF-TuSEQ. ID NO: 630 Acidovorax avenae RI N-terminus of  26TTKGHDVHGITGVNVHPKT EF-Tu REFKGKA SEQ. ID NO: 631 Acidovorax avenaeRI EF-Tu fragment 50 GSISFVDEVPMLFAGDVAR SEQ ID NO: 632EPTPIYTDLAEALKDIAQE Acidovorax spp. GLPGKDGELALK RI N-terminus of  18GITGINVHPKSREFKAKA EF-Tu SEQ. ID NO: 633 Bacillus cereusRI N-terminus of  26 TTKGHDVHGITGINVHPKS EF-Tu REFKAKA SEQ. ID NO: 634Bacillus cereus RI EF-Tu fragment 50 GITSFVDEIPMLFPKDTER SEQ ID NO:EPTPIYADVEAMLEIIKEE 635 WEAEGQLAKLAS Bacillus cereus RI N-terminus of 26 TTKGHDVHGITGVNVHPKT EF-Tu REFKGKA SEQ. ID NO: 636 Burkholderia spp.RI EF-Tu fragment 50 GSISFVDEVPMLFAGDVAR SEQ ID NO: 637EPTPIYTDLADALNMIAVE Burkholderia spp. GLEGTDGELALK RI EF-Tu fragment 50GSISFVDEVPMLFPRDVDR SEQ ID NO: 638 TPEPIFTDLADVLKLIAPV XanthomonasGIESQDGDLALR campestris RI N-terminus of  26 TTKGHDVHGITGVNVHPKS EF-TuREFKEKA SEQ. ID NO: 639 Pseudomonas spp. RI EF-Tu 50 GSISFVDEIPMLFPKDIARSEQ ID NO: 640 EPEPIYSDLTEVLKKVATT Pseudomonas spp. GMENDDKGELAM

Thionins and Thionin-Targeting Polypeptides

The polypeptide can comprise the thionin or thionin-like polypeptide.

The thionin or thionin-like polypeptide can be fused to a phloemtargeting sequence to form a fused polypeptide, the amino acid sequenceof the phloem targeting sequence comprising any one of SEQ ID NOs:641-649, or any combination thereof, for delivering the fusedpolypeptide to vascular tissue or cells and/or phloem orphloem-associated tissue or cells in the plant or plant part.

The amino acid sequence of the phloem targeting sequence can compriseSEQ ID NO: 641.

More specifically, targeting sequences useful for targeting AMPpolypeptides, such as thionins or Flg polypeptides to the vasculartissues (xylem and phloem) can be extremely useful for treating diseasesthat colonize restricted tissues involved in the transport of fluids andnutrients (e.g., water soluble nutrients, sugars, amino acids, hormones,etc.). Vascular tissues such as the xylem transport and store water andwater-soluble nutrients and the phloem cells transport sugars, proteins,amino acids, hormones and other organic molecules in plants.

Preferred vascular/phloem targeting polypeptides useful for targetingthe thionins and flagellin-associated polypeptides as described hereinare provided in Table 18.

TABLE 18 Phloem targeting polypeptides Vascular/Phloem SEQ ID NO:targeting polypeptides Phloem targeting MSTATFVDIIIAILLPPLGVFLRFGCGVpeptide EFWICLVLTLLGYIPGIIYAIYVLTK Synthetic SEQ ID NO: 641 Salt stressMGSETFLEVILAILLPPVGVFLRYGCGV induced targetingEFWICLLLTVLGYIPGIIYAIYVLVG peptide Citrus clementina SEQ ID NO: 642Hypothetical MGTATCVDIILAVILPPLGVFLKFGCKA protein CICLEEFWICLLLTILGYIPGIIYAVYVITK Citrus trifoliata SEQ ID NO: 643 HypotheticalMADEGTATCIDIILAIILPPLGVFLKFG protein CICLE CKVEFWICLLLTIFGYIPGIIYAVYAITCitrus sinensis KN SEQ ID NO: 644 Low temperatureMADGSTATCVDILLAVILPPLGVFLKFG and salt CKAEFWICLLLTILGYIPGIIYAVYAITresponsive KK protein Citrus sinensis SEQ ID NO: 645 HypotheticalFYKQKYQVQITKAVTQNPKHFFNQSSCF protein CICLE LTLNFILFHFTLFKNQSKMADGSTATCVCitrus DILLAVILPPLGVFLKFGCKAEFWICLL clementina LTILGYIPGIIYAVYAITKKSEQ ID NO: 646 Low temperature MSTATFVDIIIAILLPPLGVFLRFGCGV and saltEFWICLVLTLLGYIPGIIYAIYVLTK responsive protein Arabidopsis thalianaSEQ ID NO: 647 Cold-inducible MSTATFVDIIIAVLLPPLGVFLRFGCGV proteinEFWICLVLTLLGYIPGIIYAIYVLTK Camelina sativa SEQ ID NO: 648Low temperature MGTATCVDIIIAILLPPLGVFLRFGCGV and saltEFWICLVLTLLGYIPGILYALYVLTK responsive protein Arabidopsis lyrataSEQ ID NO: 649

A synthetic version of a phloem targeting polypeptide (SEQ ID NO: 641)is particularly useful in targeting anti-microbial polypeptides to thephloem sieve tube and companion cells.

Anti-microbial thionin polypeptides are also provided (Table 19) and areutilized with the phloem targeting sequences provided in Table 18 fortargeting the thionin sequences into the phloem tissues of citrus aswell as other plants.

The amino acid sequence of the thionin or thionin-like polypeptide cancomprise any one of SEQ ID NOs: 650-749, such as SEQ ID NO: 651.

TABLE 19 Thionin and thionin-like sequences Thionin or Thionin-likeSEQ ID NO: Sequences-Amino Acid Thionin-like proteinRTCESQSHRFKGPCSRDSNCATV Synthetic CLTEGFSGGDCRGFRRRCRCTRP SEQ ID NO: 650CVFDEK Thionin-like protein RVCQSQSHHFHGACFSHHNCAFV Citrus sinensisCRNEGFSGGKCRGVRRRCFCSKL SEQ ID NO: 651 C Thionin-like proteinKSCCKDIMARNCYNVCRIPGTPR Avena sativa PVCATTCRCKIISGNKCPKDYPKSEQ ID NO: 652 Thionin-like protein RTCESQSHRFKGPCSRDSNCATV SyntheticCLTEGFSGGDCRGFRRRCRCTRP SEQ ID NO: 653 CVFDEK Thionin-like proteinMDSRSFGLLPLLLLILLTSQMTV Citrus sinensis LQTEARLCESQSHRFHGTCVRSHSEQ ID NO: 654 NCDLVCRTEGFTGGRCRGFRRRC FCTRIC Proteinase inhibitorMKSFFGIFLLLLILFASQEIMVP se60-like protein AEGRVCQSQSHHFHGACFSHHNCCitrus paradise AFVCRNEGFSGGKCRGVRRRCFC SEQ ID NO: 655 SKLCDefensin precursor MKSFFGIFLLLLILFASQMMVPA Citrus clementinaEGRVCQSQSHHFHGACFSHHNCA SEQ ID NO: 656 FVCRNEGFSGGKCRGARRRCFCS KLCdefensin precursor MKSFFGIFLLLLILFASQEMMVP Citrus clementinaAEGRVCQSQSHHFHGACFSHHNC SEQ ID NO: 657 AFVCRNEGFSGGKCRGARRRCFC SKLCThionin-like protein MKSFFGIFLLLLILFASQMMVPA Citrus clementinaEGRVCQSQSHHFHGACFSHHNCA SEQ ID NO: 658 FVCRNEGFSGGKCRGARRRCFCS KLCThionin-like peptide MANSMRFFATVLLLALLVMATEM Nicotiana benthamianaGPMTIAEARTCESQSHRFKGPCS SEQ ID NO: 659 RDSNCATVCLTEGFSGGDCRGFR RRCFCTRPCThionin-like protein MAKSMRFFATVLLLALLVMATEM Nicotiana sylvestrisGPTTIAEARTCESQSHRFKGPCS SEQ ID NO: 660 RDSNCATVCLTEGFSGGDCRGFR RRCFCTRPCThionin-like protein MANSMRFFATVLLLTLLVMATEM Nicotiana tabaccumGPMTIAEARTCESQSHRFKGPCS SEQ ID NO: 661 RDSNCATVCLTEGFSGGDCRGFR RRCFCTRPCThionin-like protein MANSMRFFATVLLIALLVMATEM NicotianaGPMTIAEARTCESQSHRFKGPCS tomentosiformis RDSNCATVCLTEGFSGGDCRGFRSEQ ID NO: 662 RRCFCTRPC Thionin-like protein MANSMRFFATVLLIALLVTATEMNicotiana tabaccum GPMTIAEARTCESQSHRFKGPCS SEQ ID NO: 663RDSNCATVCLTEGFSGGDCRGFR RRCFCTRPC Defensin class IMANSMRFFATVLLLTLLFMATEM Nicotiana alata GPMTIAEARTCESQSHRFKGPCASEQ ID NO: 664 RDSNCATVCLTEGFSGGDCRGFR RRCFCTRPC Leaf thioninMGSIKGLKSVVICVLVLGIVLEQ Avena sativa VQVEGKSCCKDIMARNCYNVCRISEQ ID NO: 665 PGTPRPVCATTCRCKIISGNKCP KDYPKLHGDPD Leaf thioninMGSIKGLKSVVICVLVLGIVLEH Avena sativa VQVEGKSCCKDTTARNCYNVCRISEQ ID NO: 666 PGTPRPVCATTCRCKIISGNKCP KDYPKLHGDLD Thionin Class ILGLVVAQTQVDAKSCCPSTAARN Tulipa gesneriana CYNVCRFPGTPRPVCAATCGCKISEQ ID NO: 667 ITGTKCPPDYPKLGWSTFQNSDV ADKALDVVDEALHVAKEVMKEAVERCNNACSEVCTKGSYAVTA Thionin-like MERKSLGFFFFLLLILLASQEMVprotein Class I VPSEARVCESQSHKFEGACMGDH Vitis viniferaNCALVCRNEGFSGGKCKGLRRRC SEQ ID NO: 668 FCTKLC Thionin-likeMERKSLGFFFFLLLILLASQMVV protein Class I PSEARVCESQSHKFEGACMGDHNVitis vinifera CALVCRNEGFSGGKCKGLRRRCF SEQ ID NO: 669 CTKLCdefensin Ec-AMP-D1 MERSVRLFSTVLLVLLLLASEMG Citrus sinensisLRAAEARICESQSHRFKGPCVSK SEQ ID NO: 670 SNCAAVCQTEGFHGGHCRGFRRR CFCTKRCAntimicrobial LCNERPSQTWSGNCGNTAHCDKQ Protein 1 (Ah-Amp1)CQDWEKASHGACHKRENHWKCFC Aesculus hippocastanum YFNC SEQ ID NO: 671hypothetical protein  MAKNSTSPVSLFAISLIFFLLAN DCAR Dacus carotaSGSITEVDGKVCEKPSLTWSGKC SEQ ID NO: 672 GNTQHCDKQCQDWEGAKHGACHSRGGWKCFCYFEC Cysteine-rich NLCERASLTWTGNCGNTGHCDTQ antimicrobial proteinCRNWESAKHGACHKRGNWKCFCY Clitoria ternatea FNC SEQ ID NO: 673hypothetical protein MAKKSSSFCLSAIFLVLLLVANT DCAR Dacus carotaGMVREVDGALCEKPSLTWSGNCR SEQ ID NO: 674 NTQHCDKQCQSWEGAKHGACHKRGNWKCFCYHAC Thionin-like MAKKLNAVTVSAIFLVVFLIASY Bupleurum kaoiSVGAAKEAGAEGEVVFPEQLCER SEQ ID NO: 675 ASQTWSGDCKNTKNCDNQCIQWEKARHGACHKRGGKWMCFCYFDKC defensin Dm-AMP1 = ELCEKASKTWSGNCGNTGHCDNQcysteine-rich CKSWEGAAHGACHVRNGKHMCFC antimicrobial protein YFNCDahlia merckii SEQ ID NO: 676 Thionin-like MAKISVAFNAFLLLLFVLAISEIHelianthus annuus GSVKGELCEKASQTWSGTCGKTK SEQ ID NO: 677HCDDQCKSWEGAAHGACHVRDGK HMCFCYFNCSKAQKLAQDKLRAE ELAKEKIEPEKATAKP ThioninMAKNSVAFFALLLLICILTISEF Cynara cardunculus  AVVKGELCEKASKTWSGNCGNTRvar. scolymus HCDDQCKAWEGAAHGACHTRNKK SEQ ID NO: 678HMCFCYFNCPKAEKLAQDKLKAE ELARDKVEAKEVPHFKHPIEPIH HP ThioninMAKQWVSFFALAFIVFVLAISET Cynara cardunculus QTVKGELCEKASKTWSGNCGNTKvar. scolymus HCDDQCKSWEGAAHGACHVRNGK SEQ ID NO: 679HMCFCYFNSCAEADKLSEDQIEA GKLAFEKAEKLDRDVKKAVPNVD HP defensin-like proteinMAQKVNSALIFSAIFVLFLVASY 1-DCAR-like Daucus  SVTVAEGARAGAEGEVVYPEALCcarota subsp. Sativus ERASQTWTGKCQHTDHCDNQCIQ SEQ ID NO: 680WENARHGACHKRGGNWKCFCYFD HC low-molecular-weight MASSYTLMLFLCLSIFLIASTEMcysteine-rich defensin MAVEARICERRSKTWTGFCGNTR Arabidopsis lyrataGCDSQCKSWERASHGACHAQFPG SEQ ID NO: 681 FACFCYFNC Thionin-like proteinMAKSSTSYLVFLLLVLVVAISEI Parthenium ASVNGKVCEKPSKTWFGNCKDTE hysterophorusKCDKRCMEWEGAKHGACHQRESK SEQ ID NO: 682 YMCFCYFDCDP putative defensinMASSYTLMLFLCLSIFLIASTEM AMP1 protein MAVEGRICERRSKTWTGFCGNTRArabidopsis thaliana GCDSQCKRWERASHGACHAQFPG SEQ ID NO: 683 FACFCYFNCThionin-like MASSYTLLLFVCLSIFFIASTEM Eutrema salsugineumMMVEGRVCERRSKTWTGFCGNTR SEQ ID NO: 684 GCDSQCKRWERASHGACHAQFPG FACFCYFNCdefensin-like MAKLLGYLLSYALSFLTLFALLV Vitis viniferaSTEMVMLEAKVCQRPSKTWSGFC SEQ ID NO: 685 GSSKNCDRQCKNWEGAKHGACHAKFPGVACFCYFNC Knottin MAKSLSSFATFLALLCLFFLLST Corchorus olitoriusPNEMKMAEAKICEKRSQTWSGWC SEQ ID NO: 686 GNSSHCDRQCKNWENARHGSCHADGLGWACFCYFNC Knottin MEMKMAEGKICEKRSQTWSGWCG Corchorus olitoriusNSSHCDRQCKNWENARHGSCHAD SEQ ID NO: 687 GLGWACFCYFNC Thionin-like proteinMASSLKLMLFLCLSIFLIASTEM Camelina saliva MTVEGRTCERRSKTWTGFCGNTRSEQ ID NO: 688 GCDSQCRSWEGASHGACHAQFPG FACFCYFNC Thionin-like proteinMAKVVGNSAKMIVALLFLLALML Cucumis sarivus SMNEKQGVVEAKVCERRSKTWSGSEQ ID NO: 689 WCGNTKHCDRQCKNWEGATHGAC HAQFPGRACFCYFNCThionin-like protein MIDAFNYKQFSTVKGKICEKPSK Cynara cardunculusTWFGKCQDTTKCDKQCIEWEDAK var. scolymus HGACHERESKLMCFCYYNCGPPKSEQ ID NO: 690 NTPPGTPPSPP Thionin-like MASSYKLILFLCLSIFLIASFEMCapsella rubella MAVEGRICQRRSKTWTGFCGNTR SEQ ID NO: 691GCDSQCKRWERASHGACHAQFPG FACFCYFNC Thionin MMAVEGRICERRSKTWTGFCGNTArabidopsis thaliana RGCDSQCKRWERASHGACHAQFP SEQ ID NO: 692 GFACFCYFNCThionin MASSYTRLLLLCLSIFLIASTEV Brassica napus MMVEGRVCQRRSKTWTGFCGNTRSEQ ID NO: 693 GCDSQCKRWERASHGACHAQFPG FACFCYFNC Thionin-like proteinMASSYARLLLLCLSIFLIASTEV Brassica rapa MMVEGRVCQRRSKTWTGFCGNTRSEQ ID NO: 694 GCDSQCKRWERASHGACHAQFPG FACFCYFNC Thionin-like proteinMASSLKLMLFLCLSIFLIASTEM Camelina saliva MTVEGRTCERRSKTWTGFCGNTRSEQ ID NO: 695 GCDSQCRRWEHASHGACHAQFPG FACFCYFNC defensin-like proteinMASYTRLLLLCLSIFLIASTEVM Brassica napus MVEGRVCQRRSKTWTGFCGNTRGSEQ ID NO: 696 CDSQCKRWERASHGACHAQFPGF ACFCYFNC Thionin-like proteinMVMLEAKVCQRPSKTWSGFCGSS Vitis vinifera KNCDRQCKNWEGAKHGACHAKFPSEQ ID NO: 697 GVACFCYFNC Thionin-like protein MTKSFILVALLCICFILLSPTEMBrassica napus RLTLNACLKLAEAKICEKYSQTW SEQ ID NO: 698SGRCTKTSHCDRQCINWEDARHG ACHQDKHGRACFCYFNCKK Thionin-like proteinMASSYTVFLLLCLSIFLIASTEV Raphanus sativus MMVEGRVCQRRSKTWTGFCGNTRSEQ ID NO: 699 GCDSQCKRWEHASHGACHAQFPG FACFCYFNC Thionin-likeMASSYTLLLFLCLSIFLIVSTEM Arabis alpine MMVEGRICERRSKTWTGFCANTRSEQ ID NO: 700 GCDSQCKRWERASHGACHAQFPG VACFCYFNC Thionin-like proteinMAKVVGNSAKMIVAFLFLLALTL Cucumis melo SMNEKQGVVEAKVCERRSKTWSGSEQ ID NO: 701 WCGDTKHCDRQCKNWEGAKHGAC HAQFPGRACFCYFNCThionin-like protein MAASLVYRLSSVILIVLLLFIML Erythranthe guttateNNEVMVVESRLCERRSKTWTGFC SEQ ID NO: 702 GSSNNCNNQCRNWERASHGACHAQFPGFACFCYFNC Thionin-like protein MAKFQVSSTIFFALFFCFLLLASSesamum indicum NEAKICQRMSKTWSGVCLNSGNC SEQ ID NO: 703DRQCRNWERAQHGACHRRGLGFA CLCYFKC Thionin-like proteinMAKNSVAFFAFLLILFVLAISEI Eclipta prostrata GSVKGELCEKASQTWSGTCRITSSEQ ID NO: 704 HCDNQCKSWEGAAHGACHVRGGK HMCFCYFSHCAKAEKLTQDKLKAGHLVNEKSEADQKVPVTP Gamma thionin Cynara MAKNTKVSAFLFVFLFVFFLVVHcardunculus SVTAFAIRFKCFDTDMLLKVIAD var. scolymusMVVGMKGIEKVCRRRSKTWSGYC SEQ ID NO: 705 GDSKHCDQQCREWEGAEHGACHHEGLGRACFCYFNC Art v 1 precursor MAAGLLVFVLAISEIASVKGKLC AmbrosiaEKPSVTWSGKCKVKQTDKCDKRC artemisiifolia IEWEGAKHGACHKRDSKASCFCYSEQ ID NO: 706 FDCDPTKNPGPPPGAPKGKAPAP SPPSGGGGEGGGEGGGERArt v 1 precursor MAAGLLVFVLAISEIASVKGKLC AmbrosiaEKPSLTWSGKCKVKQTDKCDKRC artemi679siifolia IEWEGAKHGACHKRDSKATCFCYSEQ ID NO: 707 FDCDPTKNPGPPPGAPKGKAPAP SPPSGGGAPPPSGGEGGERThionin-like protein MAKLHSSALCFLIIFLFLLVSKE Jatropha curcasMAVTEAKLCQRRSKTWSGFCGDP SEQ ID NO: 708 GKCNRQCRNWEGASHGACHAQFPGFACFCYFKC Thionin-like protein MAKAPKSVSYFAFFFILFLLASS Nelumbo nuciferaEIQKTKKLCERRSKTWSGRCTKT SEQ ID NO: 709 QNCDKQCKDWEYAKHGACHGSWFNKKCYCYFDC Thionin-like protein MAKLLSRLSIPLIVFVFLLILLAPyrus x bretschneideri STEVAMVEARICQRRSKTWSGFC SEQ ID NO: 710ANTGNCNRQCTNWEGALHGACHA QFPGVACFCYFRC Low-molecular-weight MAKLHFPTLLCLFIFLFLLVSTE cysteine-rich protein MQVTQAKVCQRRSKTWSGFCGSTLCR78 precursor KNCDRQCKNWEGALHGACHAQFP Ricinus communi GVACFCYFKCGGERSEQ ID NO: 711 homologue of Art KLCEKPSVTWSGKCKVKQTDKCD v 1 precursorKRCIEWEGAKHGACHKRDSKASC Ambrosia FCYFDCDPTKNPGPPPGAPKGKA artemisiifoliaPAPSPPSGGGAPPPSGGEGGGD SEQ ID NO: 712 homologue of ArtKLCEKPSVTWSGNKVKQTDKCDK v 1 precursor RCIEWEGAKHGACHKRDSKASCF AmbrosiaCYFDCDPTKNPGPPPGAPKGKAP artemisiifolia APSPPSGGGAPPPSGGEGGGDGGSEQ ID NO: 713 GGRR Thionin-like protein MAKLLSHLLFYPILFLFLFIFLAPrunus mume STEVAILEARICQRRSKTWSGFC SEQ ID NO: 714GNTRNCNRQCRNWEGALRGACHA QFPGFACFCYFRC Knottin MAKTLQLFALFFIVILLANQEIPCorchorus olitorius VAEAKLCQKRSKTWTGICIKTKN SEQ ID NO: 715CDNQCKKWEKAEHGACHRQGIGF ACFCYFNQKKC Knottin MAKFVSTVALLFALFILLASFDECorchorus olitorius GMMPMAEAKVCSKRSKTWSGFCN SEQ ID NO: 716SSANCNKQCREWEDAKHGACHFE FPGFACFCYFNC Thionin-like proteinMNSKVILALLVCFLLIASNEMQG Solanum pennellii GEAKVCGRRSSTWSGLCLNTGNCSEQ ID NO: 717 NTQCIKWEHASSGACHRDGFGFA CFCYFNC Thionin-like proteinMAKLLGYHLVYPILFLFIFLLLA Fragaria vesca STEMGMLEARICQRRSKTWTGLCsubsp. Vesca ANTGNCHRQCRNWEGAQRGACHA SEQ ID NO: 718 QFPGFACFCYFNCKnottin MAKFVSVALLLALFILVASFDEG Corchorus capsularisMVPMAEAKLCSKRSKTWSGFCNS SEQ ID NO: 719 SANCNRQCREWEDAKHGACHFEFPGFACFCYFDC Thionin-like protein  MQGGEARVCERRSSTWSGPCFDTSolanum tuberosum GNCNRQCINWEHASSGACHREGI SEQ ID NO: 720 GSACFCYFNCDefensin 1.2-like MAKTLKSVQFFALFFLVILLAGS protein PDF1.2-1EMTAVEALCSKRSKTWSGPCFIT Dimocarpus longan SRCDRQCKRWENAKHGACHRSGWSEQ ID NO: 721 GFACFCYFNKC Thionin-like protein MAKAATIVTLLFAALVFFAALETCamelina sativa PTMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 722SNACKNQCINLEKARHGSCNYVF PAHKCICYFPC Thionin-like MAKFASIIAFLFAALVLFASFEAArabis alpine PTMVEAQKYCEKPSGTWSGVCGN SEQ ID NO: 723SNACNNQCINLEGARHGSCNYVF PYYRCICYFQC Thionin-like MAMSLKSVHFFALFFIVVLLANQTheobroma cacao EMPVAEAKLCQKRSKTWTGPCIK SEQ ID NO: 724TKNCDHQCRKWEKAQHGACHWQW PGFACFCYVNC Thionin-like MAKLVSPKAFFVFLFVFLLISASAmborella trichopoda EFSGSEAKLCQKRSRTWSGFCAN SEQ ID NO: 725SNNCSRQCKNLEGARFGACHRQR IGLACFCYFNC low-molecular-weightMAKSATIVTLFFAALVFFAALEA cysteine-rich 67 PMVVEAQKLCERPSGTWSGVCGNArabidopsis thaliana SNACKNQCINLEKARHGSCNYVF SEQ ID NO: 726 PAHKCICYFPCThionin-like MAKFASIITLLFAALVLFASLEA Arabis alpinePTMVEAQKLCQRPSGTWSGVCGN SEQ ID NO: 727 NGACKNQCINLEKARHGSCNYVFPYHRCICYFPC Thionin-like MAKVASIIALLFAALVLFAAFEA Brassica junceaPTMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 728 NNACKNQCINLEKARHGSCNYVFPAHKCICYFPC Thionin-like MAKFASIIALLFAALVLFAALEA Brassica oleraceaPTMVEAQKLCERPSGTWSGVCGN var. oleracea NNACKNQCINLEKARHGSCNYVFSEQ ID NO: 729 PAHKCICYFPC Thionin-like MAKPATIVTLLFAALVFFAALETCamelina sativa PTMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 730NNACKNQCINLEKARHGSCNYVF PAHKCICYFPC Thionin-like MAKSATIVTLLFAALVFFAALETCamelina sativa PTMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 731NNACKNQCINLEKARHGSCNYVF PAHKCICYFPC Thionin-like MAKFASIIAPLFAVLVLFAAFEABrassica napus PTMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 732NNACKNQCINLEKARHGSCNYVF PAHKCICYFPC Thionin-like MAKFASIITLLFAALVLFAVFEGEutrema salsugineum PTMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 733NNACKNQCINLEKARHGSCNYVF PAHKCICYFPC Cysteine-richMAKFASIIALLFAALVLFAAFEA antifungal protein PTMVEAQKLCERPSGTWSGVCGNRaphanus sativus NNACKNQCINLEKARHGSCNYVF SEQ ID NO: 734 PAHKCICYFPCThionin-like protein 1 MAKFASIVSLLFAALVLFTAFEA Raphanus sativusPAMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 735 NNACKNQCINLEKARHGSCNYVFPAHKCICYFPC Thionin-like protein 1 MNTKVILALLFCFLLVASNEMQVRaphanus sativus GEAKVCQRRSKTWSGPCINTGNC SEQ ID NO: 736SRQCKQQEDARFGACHRSGFGFA CFCYFKC Thionin-like MAKFASIIAPLFAALVLFAAFEABrassica rapa PTMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 737NNACKNQCINLEKARHGSCNYVF PAHKCICYFPC Thionin-like MNTKLILALMFCFLLIASNEMQVSolanum pennellii GEAKVCQRRSKTWSGPCINTGNC SEQ ID NO: 738SRQCKQQEDARFGACHRSGFGFA CFCYFKC Thionin-like MAKFTTTFALLFAFFILFAAFDVCitrus clementina PMAEAKVCQRRSKTWSGLCLNTG SEQ ID NO: 739NCSRQCKQQEDARFGACHRQGIG FACFCYFKC Thionin-like MAKFTSIIVLLFAALVLFAGFEABrassica rapa PTMVEAQKLCERPSGTWSGVCGN SEQ ID NO: 740NNACKNQCIRLEKARHGSCNYVF PARKCICYFPC Thionin-like MAKFASIITLLFAALVLFATFAPEutrema salsugineum TMVEAKLCERPSGTWSGVCGNNN SEQ ID NO: 741ACKSQCQRLEGARHGSCNYVFPA HKCICYFPC Thionin-like MAKFASIITLLFAALVLFATFEAEutrema salsugineum PTMVEAKLCERPSGTWSGVCGNN SEQ ID NO: 742NACKSQCQRLEGARHGSCNYVFP AHKCICYFPC Thionin-like MAKFASIIAFFFAALVLFAAFEAHeliophila  PTIVEAQKLCERPSGTWSGVCGN coronopifoliaNNACRNQCINLEKARHGSCNYVF SEQ ID NO: 743 PAHKCICYFPC Thionin-likeMAKVASIVALLFPALVIFAAFEA Brassica oleracea PTMVEAQKLCERPSGTWSGVCGNSEQ ID NO: 744 NNACKNQCIRLEKARHGSCNYVF PAHKCICYFPC Thionin-likeMSKFYTVFMFLCLALLLISSWEV Cicer arietinum EAKLCQRRSKTWSGPCIITGNCKSEQ ID NO: 745 NQCKNVEHATFGACHRQGFGFAC FCYFNCH Thionin-likeMAKSVASITTAFALIFAFFILFA Citrus clementina SFGVPMAEAKVCQRRSKTWSGPCSEQ ID NO: 746 LNTGKCSRQCKQQEYARYGACYR QGAGYACYCYFNC Thionin-likeMAKSVASITTAFALIFAFFILFA Citrus sinensis SFEVPMAEAKVCQRRSKTWSGPCSEQ ID NO: 747 LNTGKCSRHCKQQEDARYGACYR QGTGYACFCYFEC Thionin-likeMAKFTTTFALLFAFFILFAAFDV Citrus sinensis PMAEAKVCQLRSKTWSGLCLNTGSEQ ID NO: 748 NCSRQCKQQEDARFGACHRQGIG FACFCYFKC Ec-AMP-D1MERSVRLFSTVLLVLLLLASEMG Citrus sinensis LRAAEARICESQSHRFKGPCVSKSEQ ID NO: 749 SNCAAVCQTEGFHGGHCRGFRRR CFCTKRC

The polypeptide can comprise a fusion protein.

Table 20 (SEQ ID NO: 750) describes the sequences used to make atranslational fusion using the nucleotide sequence that encodes thesynthetic phloem targeting polypeptide (SEQ ID NO: 641) with a syntheticthionin polypeptide (SEQ ID NO: 650). The upper case (not bold) fontsequence identifies the phloem targeting sequence, the upper case boldfont identifies the fusion of these two peptide sequences (Table 20)that codes for the phloem targeted bioactive priming polypeptide.

TABLE 20 Translational fusion of a phloem targetingsequence with a thionin derived polypeptideTranslational fusion phloem targeting sequence withthionin polypeptide (synthetic) SEQ ID NO: 750MSTATFVDIIIAILLPPLGVFLRFGCGVEFWICLVLTLLGYIPGIIYAIYVLTKRTCESQSHRFKGPCSRDSNCATVCLTEGFSGG DCRGFRRRCRCTRPCVFDEK

Additional Modifications

In addition, polypeptides can be chemically synthesized with D-aminoacids, β2-amino acids, β3-amino acids, homo amino acids, gamma aminoacids, peptoids, N-methyl amino acids, and other non-natural amino acidmimics and derivatives.

The polypeptides may be modified by either natural processes, such asposttranslational processing, or by chemical modification techniquesthat are well known in the art. Modifications can occur anywhere in apolypeptide, including the polypeptide backbone, the amino acidside-chains and the amino or carboxyl termini. The same type ofmodification may be present in the same or varying degrees at severalsites in a polypeptide. Also, a polypeptide may contain many types ofmodifications.

Peptides may be branched, for example, as a result of ubiquitination,and they may be cyclic, with or without branching. Cyclic, branched, andbranched cyclic polypeptides may result from posttranslation naturalprocesses or may be made by synthetic methods.

Modifications include acetylation, acid addition, acylation,ADP-ribosylation, aldehyde addition, alkylamide addition, amidation,amination, biotinylation, carbamate addition, chloromethyl ketoneaddition, covalent attachment of a nucleotide or nucleotide derivative,cross-linking, cyclization, disulfide bond formation, demethylation,ester addition, formation of covalent cross-links, formation ofcysteine-cysteine disulfide bonds, formation of pyroglutamate,formylation, gamma-carboxylation, glycosylation, GPI anchor formation,hydrazide addition, hydroxyamic acid addition, hydroxylation,iodination, lipid addition, methylation, myristoylation, oxidation,PEGylation, proteolytic processing, phosphorylation, prenylation, palmitoylation, addition of a purification tag, pyroglutamyl addition,racemization, selenoylation, sulfonamide addition, sulfation,transfer-RNA mediated addition of amino acids to proteins such asarginylation, ubiquitination, and urea addition. (see, e.g., Creightonet al. (1993) Proteins—Structure and Molecular Properties, 2nd Ed., T.E. Creighton, W. H. Freeman and Company, New York; Johnson, ed. (1983)Posttranslational Covalent Modification Of Proteins, Academic Press, NewYork; Seifter et al. (1990) Meth. Enzymol., 182: 626-646; Rattan et al.(1992) Ann. N.Y. Acad. Sci., 663: 48-62; and the like).

Using known methods of protein engineering and recombinant DNAtechnology, variants may be generated to improve or alter thecharacteristics of the polypeptides described herein. Such variantsinclude deletions, insertions, inversions, repeats, duplications,extensions, and substitutions (e.g., conservative substitutions)selected according to general rules well known in the art so as havelittle effect on activity.

The polypeptide can comprise an amino acid sequence having at least 70%identity to any one of SEQ ID NOs. 1-768 wherein the polypeptide hasbioactive priming activity.

The polypeptide can comprise an amino acid sequence having at least 75%identity to any one of SEQ ID NOs. 1-768, wherein the polypeptide hasbioactive priming activity.

The polypeptide can comprise an amino acid sequence having at least 80%identity to any one of SEQ ID NOs. 1-768, wherein the polypeptide hasbioactive priming activity.

The polypeptide can comprise an amino acid sequence having at least 85%identity to any one of SEQ ID NOs. 1-768, wherein the polypeptide hasbioactive priming activity.

The polypeptide can comprise an amino acid sequence having at least 90%identity to any one of SEQ ID NOs. 1-768, wherein the polypeptide hasbioactive priming activity.

The polypeptide can comprise an amino acid sequence having at least 95%identity to any one of SEQ ID NOs. 1-768, wherein the polypeptide hasbioactive priming activity.

The polypeptide can comprise an amino acid sequence having at least 98%identity to any one of SEQ ID NOs. 1-768, wherein the polypeptide hasbioactive priming activity.

The polypeptide can comprise an amino acid sequence having at least 99%identity to any one of SEQ ID NOs. 1-768, wherein the polypeptide hasbioactive priming activity.

II. Preparation of Bioactive Priming Polypeptides

Methods and approaches are provided for cloning, genetically modifyingand expressing the bioactive priming polypeptides (for example,flagellins) and the bioactive priming polypeptides (for example,Bt.4Q7Flg22) using those methods well understood and commonly used byone of ordinary skill in the art. The methods described herein can beused with any of the bioactive priming polypeptides as described hereinand therefore include any of the flagellins, flagellin-associatedpolypeptides, thionins, harpin-like (HpaG-like), EF-Tu, PSKα or RHPPand/or any combinations thereof.

Bioactive priming polypeptides can be provided as a free polypeptide,immobilized on the surface of a particle, or impregnated on or into amatrix. Several expression systems can be used for the production offree polypeptide.

The flagellin-derived full-coding, partial coding (flagellinpolypeptides) and flagellin-associated polypeptides can be overexpressedin Bacillus strain, for example, Bacillus thuringiensis strain BT013A,in Bacillus cereus or in Bacillus subtilis. The flagellins andflagellin-derived polypeptides are cloned using an appropriateexpression vector to allow for the abundant production of thepolypeptide.

For example, in order to facilitate cloning of the target nucleotidesthat encode the bioactive priming polypeptide(s) as described herein, anE. coli compatible shuttle vector pSUPER was constructed by fusing thepBC plasmid backbone described above with the E. coli pUC57 cloningvector at compatible BamHI restriction endonuclease sites. Theresulting, pSUPER vector carries dual selection markers (ampicillinselection in E. coli and tetracycline selection in Bacillus spp).Cloning was performed by PCR amplification of target nucleotides withspecific primers synthesized with 15 bp overlapping the pSUPER insertionsite. Specific gene encoding polypeptides were fused to the pSUPERvector with In-Fusion HD Cloning Kit (Clontech). Sequence verifiedpSUPER constructs were amplified using the pBC suitable backbone Reverseand Forward primers. The resulting PCR products were self-ligated togenerate the pBC plasmid that was used to transform the B30 donorBacillus spp. strain. The final construct was verified to be completelyintrageneric by Sanger sequencing.

The bioactive priming polypeptides/peptides as described herein areproduced in large amounts for field and grower applications by using afree expression system that can utilize a Bacillus subtilis and/orBacillus thuringiensis strain as the designated heterologous expressionstrain. The base expression plasmid designated pFEe4B consists of an E.coli section (=e) and a Bacillus section (=pFE). The e section wasderived from pUC19 and enables selection and amplification of the vectorin E. coli for cloning purposes. It comprises the beta-lactamase gene(bla) conferring resistance to beta-lactam antibiotics such asampicillin and other penicillin derivatives, as well as an E. coliorigin of replication allowing vector multiplication. The pFE sectionprovides selection and plasmid amplification in Bacillus spp. and drivesexpression of the heterologous polypeptide/peptide of interest. As suchit contains a gene conferring resistance to tetracycline (tetL), as wellas the gene for a replication protein (repU) responsible for amplifyingthe plasmid in Bacillus spp., both of which were derived from the nativeBacillus cereus plasmid pBC16. The expression cassette of pFEe4Bcontains a secretion signal (amyQ), a cloning site and a terminator(rspD), the former resulting in secretion of the expressedprotein/peptide from the host strain cells into the surrounding medium,and the latter preventing transcription beyond the open reading frame ofinterest. Expression in pFEe4B is driven by a modified autoinduciblepromoter, which initiates expression once the culture reaches asufficient optical density. In the pFEe4b expression system, expressionis controlled by an IPTG-inducible promoter sequence from Bacillussubtilis. This promoter consists of a modified constitutive promotercombined with the E. coli lac repressor (lacI) and a ribosome bindingsite. Thus, expression from pFEe4B-encoded polypeptides/peptides dependson the presence of suitable induction agents such as isopropylbeta-D-1-thiogalactopyranoside (IPTG). However other pFe systems usefulfor expression of the polypeptides as described herein do not rely onsuch induction systems for their expression. The pFEe4 plasmid furtherharbors the E. coli lacI gene under control of the Bacilluslicheniformis penicillase promoter to prevent expression ofpolypeptide/peptide as described herein in absence of any inductionagent.

Other commercially available expression vectors, for example, any ofthose derived from Bacillus subtilis, can also be useful. Otherexpression vectors were selected for producing the recombinant bioactivepriming polypeptides due to the following desired criteria: therecombinant microorganism is non-pathogenic and is considered asgenerally regarded as safe (GRAS) organisms, it has no significant biasin codon usage and it is capable of secreting extracellular proteinsdirectly into the culture medium providing for a cell free version(s) ofthe bioactive priming polypeptides.

Other expression systems common in the art can be utilized to expressbioactive priming polypeptides in a similar manner.

The bioactive priming polypeptides as described herein can be producedand purified either by the use of a protein tag(s) using affinitypurification or by using column protease cleavage methods which releasethe un-tagged polypeptide(s). Methods of using this approach to makefree versions of the bioactive priming polypeptides are commonly knownand understood by one of ordinary skill in the art.

Protein tags usually comprise a relatively small sequence of amino acidsincorporated into a translated polypeptide, basically providing amolecular tether for the bioactive priming polypeptide of interest. Theyare commonly used to aid in the expression and purification ofrecombinant polypeptides. The polyhistidine (His) tag was selected forthe purposes of affinity purification of the bioactive primingpolypeptides as described. A His tag can be fused to either the N- orC-terminus of a polypeptide. His tags are frequently combined with othertags for dual-labeling. Tags for the bioactive priming polypeptides canbe useful to affinity purify them. The tags can also be cleaved off ofthe bioactive priming polypeptides using specific proteases andcolumn-specific protease cleavage methods to release the purifiedun-tagged bioactive priming polypeptide or full-length precursor proteinof interest. These methods are also common and well known to one ofordinary skill in the art. Other tags that can be utilized are known inthe art, and include FLAG tags, antibody epitopes, streptavidin/biotin,among other purification tools. Another useful tag is a glutathioneS-transferase (GST) tag.

Protein tags can be provided within the plasmid to produce thepolypeptide. Ideally, the plasmid comprises, alongside the sequenceencoding the polypeptide of interest, a secretion signal (e.g., the amyEor amyQ secretion signal) to promote secretion, and a protein tag (e.g.,glutathione S transferase) to enhance the stability of the polypeptide,thereby enhancing production and stability. In preferred cases, theprotein tag (e.g., GST) is linked to the polypeptide using a linkersequence comprising a consensus cleavage sequence. This can allow theaddition of a targeted kinase that can cleave the tag and release thepurified, isolated polypeptide. A suitable consensus cleavage sequencecan comprise an enterokinase cleavage sequence (SEQ ID NO: 772), whichcan be cleaved by simple application of a bovine enterokinase, forexample.

Therefore, a method is provided for producing a polypeptide comprisingproducing a fusion protein comprising any polypeptide described hereinand an Enterokinase (EK) cleavage site via fermentation, the EK cleavagesite serving to enhance activity and stability of the polypeptide. Thefusion protein encoded by the plasmid can further comprise a protein tag(e.g., a poly-histidine (His) tag, a FLAG tag, an antibody epitope,streptavidin/biotin, glutathione S-transferase (GST), or any combinationthereof), wherein the enterokinase cleavage site comprises a linkingregion connecting the polypeptide and the protein tag. The fusionprotein can also comprise a secretion signal. The secretion signal cancomprise an amyE or amyQ secretion signal (e.g., SEQ ID NO: 769), or itcan comprise any one of SEQ ID NOs 563-570 as described above. Thepolypeptide comprising the enterokinase (EK) cleavage site can be morestable and produced in higher yields using fermentation than apolypeptide lacking the enterokinase (EK) cleavage site. When desired,an enterokinase (e.g., a bovine enterokinase) can be applied to thefusion protein to activate (e.g., isolate) the polypeptide of interest.The enterokinase can be applied on-site to enable maximum stability ofthe bioactive priming polypeptide prior to administration.

The bioactive priming polypeptides can be provided in a synthetic formusing commercially available peptide synthesis technologies to producehigh purity polypeptides. Synthetic production of the bioactive primingpolypeptides utilizes general solid-phase peptide synthesismethodologies that are well known to one of ordinary skill in the art.Chemical synthesis methodologies include: a stepwise assembly ofpeptides from amino acid precursors, whereby peptide elongation proceedsvia a coupling reaction between amino acids, followed by the removal ofa reversible protecting group. Solid phase peptide synthesis is used toadd a covalent attachment step that links the nascent peptide chain toan insoluble polymeric support whereby the anchored peptide can beextended by a series of cycles. These extension reactions are driven tocompletion and then the synthesized polypeptide is removed from thesolid support by filtration and washing steps. MS and HPLC analyses areperformed after the completion of synthesis and purification.

Any of the bioactive priming polypeptides as described herein forflagellin-associated polypeptides (Tables 1-5), harpin-like (HpaG-like)polypeptides (Table 10 and 11), phytosulfokine (PSKα) polypeptides(Table 12), RHPP (Table 13-15), elongation factor Tu (EF-Tupolypeptides) (Tables 16 and 17), thionin and thionin-like polypeptides(Table 19) can be provided in synthetic forms.

Additionally, such methods can be used for making and using conservedassistance sequences preferably named signature (SEQ ID NOs: 542-548),signal anchor sorting (SEQ ID NOs: 549-562) and secretion (SEQ ID NOs:563-570) sequences.

Retro inverso can also be made synthetically or chemically manufactured.Synthetic polypeptides produced in the all-D confirmation are preparedby replacing all the L-amino acid residues with their D-enantiomersresulting in a reversed or retro-all-D-isomer Flg polypeptide. Solidphase synthesis is used to prepare the retro-inverso versions of the Flgpolypeptide(s). After synthesis and purification of the retro-inversopolypeptide(s), the amino acid composition is confirmed using massspectrometry of the Flg polypeptide(s). The purity of the retro-inversopolypeptide(s) is then confirmed at a level greater or equal to 95%using HPLC analysis. The retro-inverso versions of the Flgpolypeptide(s) are further characterized using HPLC retention time,relative molecular mass and amino acid composition values (IC50 μM).Retro inverso production using recombinant DNA technology generallyinvolves the use of non-ribosomal protein synthesis mechanisms.

Retro-inverso synthetic Flg bioactive priming polypeptides prepared bysolid phase synthesis are tested for their capacity to bind to the FLS2or alternative FLS receptors, for example, FLS3 also found in plants.Competitive ELISA experiments are used to confirm the binding affinitiesof retro inverso Flg-associated polypeptides to plant FLS receptors.

Recombinant Bacteria that Express Bioactive Priming Polypeptides

A recombinant microorganism that expresses or overexpresses apolypeptide is also provided. The polypeptide comprises the polypeptidesas described above for the composition. For example, the polypeptide cancomprise: the flagellin or flagellin-associated polypeptide of (a); orthe mutant flagellin or flagellin-associated polypeptide of (b); or themutant flagellin or flagellin-associated polypeptide of (c); or theharpin or harpin-like polypeptide of (g); or the RHPP of (i); or the KTIpolypeptide of (j); or the EF-Tu polypeptide of (l); or the fusionpolypeptide of (n); or the PSK polypeptide of (o); or the thionin orthionin-like polypeptide of (q).

The polypeptide can be overexpressed by the microorganism. Therecombinant microorganism can comprise a microorganism that is capableof making recombinant bioactive priming polypeptides or their precursorsin an effective manner. The preferred microorganism would be from thegenus Bacillus, a bacterium of the genus Paenibacillus, a fungus of thegenus Penicillium, a bacterium of the genus Glomus, a bacterium of thegenus Pseudomonas, a bacterium of the genus Arthrobacter, a bacterium ofthe genus Paracoccus, a bacterium of the genus Rhizobium, a bacterium ofthe genus Bradyrhizobium, a bacterium of the genus Azosprillium, abacterium of the genus Enterobacter, a bacterium of the genusEscherichia, or any combination thereof.

The recombinant microorganism can comprise a bacterium of the genusBacillus, a bacterium of the genus Paenibacillus, or any combinationthereof.

For example, the microorganism can comprise Bacillus mycoides, Bacilluspseudomycoides, Bacillus cereus, Bacillus thuringiensis, Bacillusmegaterium, Bacillus subtilis, Bacillus firmus, Bacillus aryabhattai,Bacillus amyloliquefaciens, Bacillus licheniformis, Bacillus circulans,Bacillus flexus, Bacillus nealsonii, Bacillus pumulis, Paenibacillusgenus bacterium or a combination thereof.

Methods and approaches are commonly used by one of ordinary skill in theart to determine and verify the genus and species of the bacteria. Acommon method provides chromosomal DNA isolated from the bacteria withPCR amplification of the 16s rRNA region using universal primers(ACTCCTACGGGAGGCAGCAGT) and (GGGTTGCGCTCGTTG/AC). The PCR amplicons arethen purified and sequenced for correct identification of theappropriate bacterial strain, for example a specific strain in thegenera of Bacillus.

Sample protocols are generally known to one in the art for thepreparation of chromosomal DNA, transformation of the DNA of genesencoding the polypeptides using a plasmid, producing the polypeptides ina host bacterium, for example, a Bacillus strain.

The Bacillus strains provided can produce any bioactive primingpolypeptide as described herein or a combination thereof. For example,the strain can comprise:

(a) Bacillus aryabhattai CAP53 (NRRL No. B-50819),(b) Bacillus aryabhattai CAP56 (NRRL No. B-50817),(c) Bacillus flexus BT054 (NRRL No. B-50816),(d) Paracoccus kondratievae NC35 (NRRL No. B-50820),(e) Bacillus mycoides BT155 (NRRL No. B-50921),(f) Enterobacter cloacae CAP12 (NRRL No. B-50822),(g) Bacillus nealsonii BOBA57 (NRRL No. NRRL B-50821),(h) Bacillus mycoides EE118 (NRRL No. B-50918),(i) Bacillus subtilis EE148 (NRRL No. B-50927),(j) Alcaligenes faecalis EE107 (NRRL No. B-50920),(k) Bacillus mycoides EE141 (NRRL NO. B-50916),(l) Bacillus mycoides BT46-3 (NRRL No. B-50922),(m) Bacillus cereus family member EE128 (NRRL No. B-50917),(n) Paenibacillus massiliensis BT23 (NRRL No. B-50923),(o) Bacillus cereus family member EE349 (NRRL No. B-50928),(p) Bacillus subtilis EE218 (NRRL No. B-50926),(q) Bacillus megaterium EE281 (NRRL No. B-50925),(r) Bacillus cereus family member EE-B00377 (NRRL B-67119);(s) Bacillus pseudomycoides EE-B00366 (NRRL B-67120),(t) Bacillus mycoides EE-B00363 (NRRL B-67121),(u) Bacillus pumilus EE-B00143 (NRRL B-67123),(v) Bacillus thuringiensis EE-B00184 (NRRL B-67122),(w) Bacillus mycoides EE116 (NRRL No. B-50919),(x) Bacillus cereus family member EE417 (NRRL No. B-50974),(y) Bacillus subtilis EE442 (NRRL No. B-50975),(z) Bacillus subtilis EE443 (NRRL No. B-50976),(aa) Bacillus cereus family member EE444 (NRRL No. B-50977),(bb) Bacillus subtilis EE405 (NRRL No. B-50978),(cc) Bacillus cereus family member EE439 (NRRL No. B-50979),(dd) Bacillus megaterium EE385 (NRRL No. B-50980),(ee) Bacillus cereus family member EE387 (NRRL No. B-50981),(ff) Bacillus circulans EE388 (NRRL No. B-50982),(gg) Bacillus thuringiensis EE319 (NRRL No. B-50983),(hh) Bacillus cereus family member EE377 (NRRL No. B-67119),(ii) Bacillus mycoides EE363 (NRRL No. B-67121),(jj) Bacillus pseudomycoides EE366 (NRRL No. B-67120);(kk) Bacillus thuringiensis BT013A (NRRL No. B-50924);

or any combination thereof. Each of these strains has been depositedwith the United States Department of Agriculture (USDA) AgriculturalResearch Service (ARS), having the address 1815 North University Street,Peoria, Ill. 61604 U.S.A., and are identified by the NRRL depositnumbers provided in parentheses. Strains (a)-(d), (f), and (g) weredeposited on Mar. 11, 2013. Strains (e), (h)-(q), (w), and (kk) weredeposited on Mar. 10, 2014. Strains (x)-(ff) were deposited on Sep. 10,2014. Strain (gg) was deposited on Sep. 17, 2014. Strains (r)-(v), (hh),(ii), and (jj) were deposited on Aug. 19, 2015. Bacillus thuringiensisBT013A is also known as Bacillus thuringiensis 4Q7.

The isolation and characterization of these strains are described in theExamples found within International Publication No: WO/2017/161091,incorporated herein by reference in its entirety. For ease ofidentification of the organism, International Publication No:WO/2017/161091 A1 also provides the partial 16S ribosomal RNA sequencesfor each of these strains in a sequence list and in Table 17.

Any of the recombinant microorganisms can be used to overexpress abioactive priming polypeptide as described herein for aflagellin-associated polypeptide (Tables 1-5), a harpin or harpin-like(HpaG-like) polypeptide (Table 10 or 11), a phytosulfokine (PSKα)polypeptide (Table 12), RHPP (Table 13-15), an EF-Tu polypeptide (Table16-17, and a thionin or thionin-like polypeptide (Table 19).

The recombinant microorganism can comprise a mixture of two or more ofany of the recombinant microorganisms described herein.

The recombinant microorganism can be inactivated. Inactivation resultsin microorganisms that are unable to reproduce. Inactivation ofmicroorganisms can be advantageous, for example because it allows fordelivery of the microorganism to a plant or a plant growth medium whilereducing or eliminating any detrimental effects that the livemicroorganism may have on a plant or on the environment. The recombinantmicroorganism can be inactivated by any physical or chemical means,e.g., by heat treatment, gamma irradiation, x-ray irradiation, UV-Airradiation, UV-B irradiation, or treatment with a solvent such asglutaraldehyde, formaldehyde, hydrogen peroxide, acetic acid, bleach,chloroform, or phenol, or any combination thereof.

III. Compositions

A composition is provided for bioactive priming of a plant or a plantpart to increase growth, yield, health, longevity, productivity, and/orvigor of a plant or a plant part and/or decrease abiotic stress in theplant or the plant part and/or protect the plant or the plant part fromdisease, insects and/or nematodes, and/or increase the innate immuneresponse of the plant or the plant part and/or change plantarchitecture. The composition comprises either: the polypeptide asdescribed herein or any combination thereof, and an agrochemical or acarrier; or any combination of the polypeptides as described herein.

The composition can consist essentially of the bioactive primingpolypeptides or polypeptides as described herein.

The composition can comprise a majority of the bioactive primingpolypeptides with the remainder of the composition being agrochemicalsor carriers. More specifically, the composition can comprise from about0.00001% to about 95% of the polypeptides, from about 0.1 to about 80wt. % of the agrochemicals, and from about 5 to about 50 wt. % carrierbased on the total weight of the composition. Alternatively, thecomposition can comprise from about 0.01 to about 5 wt. % of thepolypeptides, from about 0.2 to about 70 wt. % of the agrochemicals, andfrom about 10 to about 30 wt. % carrier based on the total weight of thecomposition, or the composition can comprise from about 0.05 wt. % toabout 1 wt. % of the polypeptides, from about 30 to about 60 wt. % ofthe agrochemicals, and from about 40 to about 69 wt. % carrier based onthe total weight of the composition. Alternatively, the composition cancomprise any detectable amount of the polypeptides, and from about 0.1to about 80 wt. % of the agrochemicals and from about 5 to about 50 wt.% of the carrier, based on the total weight of the composition.

The composition can include either an agrochemical or a carrier which isassociated with the polypeptide in nature.

The agrochemical can be non-naturally occurring in combination with thepolypeptide.

The agrochemical can include, but is not limited to, a preservative, abuffering agent, a wetting agent, a surfactant, a coating agent, amonosaccharide, a polysaccharide, an abrading agent, a pesticide, aninsecticide, an herbicide, a nematicide, a bacteriocide, a fungicide, amiticide, a fertilizer, a biostimulant, a colorant, a humectant, anosmoprotectant, an antibiotic, an amino acid, a biological controlagent, or a combination thereof.

When the composition includes an amino acid, the amino acid can beprovided separately from the amino acids that comprise the polypeptide.For example, an isolated amino acid can be used. Suitable amino acidsinclude any natural or unnatural amino acids. For example, thecomposition can comprise cysteine.

The agrochemical can comprise an acid such as an acid that is presentfrom chemical synthesis of any polypeptide described herein. Forexample, hydrochloric acid, acetic acid, or trifluoroacetic acid can bepresent if the polypeptide is synthesized such as by fermentation.

When the agrochemical is an acid, it can comprise from about 0.001 toabout 30 wt. %, from about 0.01 to about 20 wt. %, or from about 0.1 toabout 5 wt. % of the total weight of the composition.

Unless otherwise specified, each agrochemical can comprise from about0.1 to about 60 wt. %, from about 0.5 to about 50 wt. %, or from about10 to about 30 wt. % of the total weight of the composition.

When the composition includes a preservative, the preservative cancomprise those based on dichlorophene and benzylalcohol hemi formal(PROXEL from ICI or ACTICIDE RS from Thor Chemie and KATHON MK from DowChemical) and isothiazolinone derivatives such as alkylisothiazolinonesand benzisothiazolinones (ACTICIDE MBS from Thor Chemie). As furtherexamples, suitable preservatives include MIT(2-methyl-4-isothiazolin-3-one), BIT (1,2-benzisothiazolin-3-one, whichcan be obtained from Avecia, Inc. as PROXEL GXL as a solution in sodiumhydroxide and dipropylene glycol),5-chloro-2-(4-chlorobenzyl)-3(2H)-isothiazolone,5-chloro-2-methyl-2H-isothiazol-3-one,5-chloro-2-methyl-2H-isothiazol-3-one,5-chloro-2-methyl-2H-isothiazol-3-one-hydrochloride,4,5-dichloro-2-cyclohexyl-4-isothiazolin-3-one,4,5-dichloro-2-octyl-2H-isothiazol-3-one, 2-methyl-2H-isothiazol-3-one,2-methyl-2H-isothiazol-3-one-calcium chloride complex,2-octyl-2H-isothiazol-3-one, benzyl alcohol hem iformal, or anycombination thereof.

When the composition includes a buffering agent, the buffering agent cancomprise potassium, phosphoric acid, a phosphate salt, citric acid, acitrate salt, a sulfate salt, MOPS, or HEPES. The buffering agent canstabilize the polypeptide in the composition.

When the composition includes a wetting agent, the wetting agent cancomprise organosilicones, polyoxyethoxylates, polysorbates,polyethyleneglycol and derivatives thereof, ethoxylates, crop oils, andpolysaccharides.

When the composition includes a surfactant, the surfactant can comprisea heavy petroleum oil, a heavy petroleum distillate, a polyol fatty acidester, a polyethoxylated fatty acid ester, an aryl alkyl polyoxyethyleneglycol, a polyoxyethylenepolyoxypropylene monobutyl ether, an alkylamine acetate, an alkyl aryl sulfonate, a polyhydric alcohol, an alkylphosphate, an alcohol ethoxylate, an alkylphenol ethoxylate, analkyphenol ethoxylate, an alkoxylated polyol, an alky polyethoxy ether,an alkylpolyoxethylene glycerol, ethoxylated and soybean oilderivatives, an organosilicone-based surfactant or any combinationthereof. Surfactants can be included in a range of compositionsincluding those for foliar use.

When the composition includes a coating agent, the coating agent cancomprise a tackifier, polymers, filling agents, or bulking agents.

The tackifier can include, but is not limited to, carboxymethylcelluloseand natural and synthetic polymers in the form of powders, granules, orlatexes, such as gum Arabic, chitin, polyvinyl alcohol and polyvinylacetate, as well as natural phospholipids, such as cephalins andlecithins, and synthetic phospholipids. Tackifiers include thosecomposed preferably of an adhesive polymer that can be natural orsynthetic without phytotoxic effect on the seed to be coated. Additionaltackifiers that can be included, either alone or in combination,include, for example, polyesters, polyether esters, polyanhydrides,polyester urethanes, polyester amides; polyvinyl acetates; polyvinylacetate copolymers; polyvinyl alcohols and tylose; polyvinyl alcoholcopolymers; polyvinylpyrolidones; polysaccharides, including starches,modified starches and starch derivatives, dextrins, maltodextrins,alginates, chitosanes and celluloses, cellulose esters, cellulose ethersand cellulose ether esters including ethylcelluloses, methylcelluloses,hydroxymethylcelluloses, hydroxypropylcelluloses andcarboxymethylcellulose; fats; oils; proteins, including casein, gelatinand zeins; gum arabics; shellacs; vinylidene chloride and vinylidenechloride copolymers; lignosulfonates, in particular calciumlignosulfonates; polyacrylates, polymethacrylates and acryliccopolymers; polyvinylacrylates; polyethylene oxide; polybutenes,polyisobutenes, polystyrene, polybutadiene, polyethyleneamines,polyethylenam ides; acrylamide polymers and copolymers; polyhydroxyethylacrylate, methylacrylamide monomers; and polychloroprene, or anycombination thereof. Tackifiers can be used in a range of compositionsincluding those for seed treatment.

When the composition includes an abrading agent, the abrading agent cancomprise talc, graphite, or a combination of both.

A humectant is a hygroscopic substance that assists with the retentionof moisture. When the composition includes a humectant, the humectantcan comprise: glycerol, glycerin, a glycerol derivative (e.g. glycerolmonosterate, glycerol triacetate, triacetin, propylene glycol, hexyleneglycol, or butylene glycol), triethylene glycol, tripolypropyleneglycol, glyceryl triacetate, sucrose, tagatose, a sugar alcohol or asugar polyol (e.g glycerol, sorbitol, xylitol, mannitol, or mantitol), apolymeric polyol (e.g. polydextrose, a collagen, an aloe or an aloe veragel), or an alpha hydroxy acid (e.g. lactic acid, honey, molasses,quillaia, sodium hexametaphosphate, lithium chloride or urea). Synthetichumectants can also comprise: butylene glycol, and tremella extract.

When the composition includes a pesticide, the pesticide can comprise aninsecticide, a herbicide, a fungicide, a bacteriocide, a nematicide, amiticide, or any combination thereof.

When the composition includes an insecticide, the insecticide cancomprise clothianidin, imidacloprid, an organophosphate, a carbamate, apyrethroid, an acaricide, an alkyl phthalate, boric acid, a borate, afluoride, sulfur, a haloaromatic substituted urea, a hydrocarbon ester,a biologically-based insecticide, or any combination thereof. Forexample, the insecticide can comprise clothianidin or imidacloprid.

The agrochemical can comprise an herbicide. The herbicide can comprise2,4-D, 2,4-DB, acetochlor, acifluorfen, alachlor, ametryn, atrazine,aminopyralid, benefin, bensulfuron, bensulfuron methyl bensulide,bentazon, bispyribac sodium, bromacil, bromoxynil, butylate,carfentrazone, chlorimuron, 2-chlorophenoxy acetic acid, chlorsulfuron,chlorimuron ethyl, clethodim, clomazone, clopyralid, cloransulam,CMPP-P-DMA, cycloate, DCPA, desmedipham, dicamba, dichlobenil, diclofop,2,4-dichlorophenol, dichlorophenoxyacetic acid, dichlorprop,dichlorprop-P, diclosulam, diflufenzopyr, dimethenamid, dimethyl aminesalt of 2,4-dichlorophenoxyacetic acid, diquat, diuron, DSMA, endothall,EPTC, ethalfluralin, ethofumesate, fenoxaprop, fluazifop-P,flucarbazone, flufenacet, flumetsulam, flumiclorac, flumioxazin,fluometuron, fluroxypyr, fluorxypyr 1-methyleptylester, fomesafen,fomesafen sodium salt, foramsulfuron, glufosinate, glufosinate-ammonium,glyphosate, halosulfuron, halosulfuron-methyl, hexazinone,2-hydroxyphenoxy acetic acid, 4-hydroxyphenoxy acetic acid,imazamethabenz, imazamox, imazapic, imazaquin, imazethapyr, isoxaben,isoxaflutole, lactofen, linuron, mazapyr, MCPA, MCPB, mecoprop,mecoprop-P, mesotrione, metolachlor-s, metribuzin, metsulfuron,metsulfuron-methyl, molinate, MSMA, napropamide, naptalam, nicosulfuron,norflurazon, oryzalin, oxadiazon, oxyfluorfen, paraquat, pelargonicacid, pendimethalin, phenmedipham, picloram, prim isulfuron, prodiamine,prometryn, pronamide, propanil, prosulfuron, pyrazon, pyrithiobac,pyroxasulfone,quinclorac, quizalofop, rimsulfuron, sethoxydim, siduron,simazine, sulfentrazone, sulfometuron, sulfosulfuron, tebuthiuron,terbacil, thiazopyr, thifensulfuron, thifensulfuron-methyl, thiobencarb,tralkoxydim, triallate, triasulfuron, tribenuron, tribernuron-methyl,triclopyr, trifluralin, triflusulfuron, or any combination thereof.

When the composition includes a nematicide, the nematicide can compriseBacillus firmus, fluopyram, antibiotic nematicides such as abamectin;carbamate nematicides such as acetoprole, Bacillus chitonosporus,chloropicrin, benclothiaz, benomyl, Burholderia cepacia, carbofuran,carbosulfan, and cleothocard; dazomet, DBCP, DCIP, alanycarb, aldicarb,aldoxycarb, oxamyl, diamidafos, fenamiphos, fosthietan, phosphamidon,cadusafos, chlorpyrifos, diclofenthion, dimethoate, ethoprophos,fensulfothion, fostiazate, harpins, heterophos, imicyafos, isamidofos,isazofos, methomyl, mecarphon, Myrothecium verrucaria, Paecilomyceslilacinus, Pasteuria nishizawae (including spores thereof), phorate,phosphocarb, terbufos, thionazin, triazophos, tioxazafen, dazomet,1,2-dicloropropane, 1,3-dichloropropene, furfural, iodomethane, metam,methyl bromide, methyl isothiocyanate, xylenol, or any combinationthereof. For example, the nematicide can comprise Bacillus firmus straini-2580, Pasteuria nishizawae (including spores thereof), or fluopyram.

When the composition includes a bacteriocide, the bacteriocide cancomprise streptomycin, penicillins, tetracyclines, oxytetracycline,kasugamycin, ampicillin, oxolinic acid, chlorotetracycline, copperoxide, or any combination thereof. For example, the bacteriocide cancomprise oxytetracycline.

Biological control agents are broadly defined as microorganisms that canbe used instead of synthetic pesticides or fertilizers. When thecomposition includes a biological control agent, the biological controlagent can comprise Bacillus thuringiensis, Bacillus megaterium, Bacillusmycoides isolate J, Bacillus methylotrophicus, Bacillus vallismortis,Chromobacterium subtsugae, Delftia acidovorans, Streptomyces lydicus,Streptomyces colombiensis, Streptomyces galbus K61, Penicillium bilaii,a lipopeptide-producing Bacillus subtilis strain, alipopeptide-producing Bacillus amyloliquefaciens strain, a Bacillusfirmus strain or a Bacillus pumilus strain.

The agrochemical can include a fungicide. The fungicide can comprisealdimorph, ampropylfos, ampropylfos potassium, andoprim, anilazine,azaconazole, azoxystrobin, benalaxyl, benodanil, benomyl, benzamacril,benzamacryl-isobutyl, benzovindflupyr, bialaphos, binapacryl, biphenyl,bitertanol, blasticidin-S, boscalid, bromuconazole, bupirimate,buthiobate, calcium polysulphide, capsimycin, captafol, captan,carbendazim, carvon, quinomethionate, chlobenthiazone, chlorfenazole,chloroneb, chloropicrin, chlorothalonil, chlozolinate, clozylacon,cufraneb, cymoxanil, cyproconazole, cyprodinil, cyprofuram, debacarb,dichlorophen, diclobutrazole, diclofluanid, diclomezine, dicloran,diethofencarb, dimethirimol, dimethomorph, dimoxystrobin, diniconazole,diniconazole-M, dinocap, diphenylamine, dipyrithione, ditalimfos,dithianon, dodemorph, dodine, drazoxolon, edifenphos, epoxiconazole,etaconazole, ethirimol, etridiazole, famoxadon, fenapanil, fenarimol,fenbuconazole, fenfuram, fenitropan, fenpiclonil, fenpropidin,fenpropimorph, fentin acetate, fentin hydroxide, ferbam, ferimzone,fluazinam, fludioxonil, flumetover, fluoromide, fluoxastrobinfluquinconazole, flurprimidol, flusilazole, flusulfamide, flutolanil,flutriafol, folpet, fosetyl-aluminium, fosetyl-sodium, fthalide,fuberidazole, furalaxyl, furametpyr, furcarbonil, furconazole,furconazole-cis, furmecyclox, guazatine, hexachlorobenzene,hexaconazole, hymexazole, imazalil, imibenconazole, iminoctadine,iminoctadine albesilate, iminoctadine triacetate, iodocarb, iprobenfos(IBP), iprodione, irumamycin, isoprothiolane, isovaledione, kasugamycin,kresoxim-methyl, copper preparations, such as: copper hydroxide, coppernaphthenate, copper oxychloride, copper sulphate, copper oxide,oxine-copper and Bordeaux mixture, mancopper, mancozeb, maneb,meferimzone, mepanipyrim, mepronil, metconazole, metalzxyl,methasulfocarb, methfuroxam, metiram, metomeclam, metsulfovax,mildiomycin, myclobutanil, myclozolin, nickel dimethyldithiocarbamate,nitrothal-isopropyl, nuarimol, ofurace, oxadixyl, oxamocarb, oxolinicacid, oxycarboxim, oxyfenthiin, paclobutrazole, pefurazoate,penconazole, pencycuron, phosdiphen, picoxystrobin, pimaricin,piperalin, polyoxin, polyoxorim, probenazole, prochloraz, procymidone,propamocarb, propanosine-sodium, propiconazole, propineb,prothiocinazole, pyrazophos, pyrifenox, pyrimethanil, pyroquilon,pyroxyfur, quinconazole, quintozene (PCNB), a strobilurin, sulphur andsulphur preparations, tebuconazole, tecloftalam, tecnazene, tetcyclasis,tetraconazole, thiabendazole, thicyofen, thifluzamide,thiophanate-methyl, tioxymid, tolclofos-methyl, tolylfluanid,triadimefon, triadimenol, triazbutil, a triazole, triazoxide,trichlamide, tricyclazole, triclopyr, tridemorph, trifloxystrobin,triflumizole, triforine, uniconazole, validamycin A, vinclozolin,viniconazole, zarilamide, zineb, ziram and also Dagger G, OK-8705,OK-8801,a-(1,1-dimethylethyl)-(3-(2-phenoxyethyl)-1H-1,2,4-triazole-1-ethanol,a-(2,4-dichlorophenyl)-[3-fluoro-3-propyl-1H-1,2,4-triazole-1-ethanol,a-(2,4-dichlorophenyl)-[3-methoxy-a-methyl-1H-1,2,4-triazole-1-ethanol,a-(5-methyl-1,3-dioxan-5-yl)-[3-[[4-(trifluoromethyl)-phenyl]-methylene]-1H-1,2,4-triazole-1-ethanol,(5RS,6RS)-6-hydroxy-2,2,7,7-tetramethyl-5-(1H-1,2,4-triazol-1-yl)-3-octanone,(E)-a-(methoxyimino)-N-methyl-2-phenoxy-phenylacetamide,1-isopropyl{2-methyl-1-[[[1-(4-methylphenyl)-ethyl]-amino]-carbonyl]-propyl}carbamate,1-(2,4-dichlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-ethanone-O-(phenylmethyl)-oxime, 1-(2-methyl-1-naphthalenyl)-1H-pyrrole-2,5-dione,1-(3,5-dichlorophenyl)-3-(2-propenyl)-2,5-pyrrolidindione,1-[(diiodomethyl)-sulphonyl]-4-methyl-benzene,1-[[2-(2,4-dichlorophenyl)-1, 3-dioxolan-2-yl]-methyl]-1H-imidazole,1-[[2-(4-chlorophenyl)-3-phenyloxiranyl]-methyl]-1H-1,2,4-triazole,1-[1-[2-[(2,4-dichlorophenyl)-methoxy]-phenyl]-ethenyl]-1H-imidazole,1-methyl-5-nonyl-2-(phenylmethyl)-3-pyrrolidinole,2′,6′-dibromo-2-methyl-4′-trifluoromethoxy-4′-trifluoro-methyl-1,3-thiazole-carboxanilide,2,2-dichloro-N-[1-(4-chlorophenyl)-ethyl]-1-ethyl-3-methyl-cyclopropanecarboxamide,2,6-dichloro-5-(methylthio)-4-pyrimidinyl-thiocyanate,2,6-dichloro-N-(4-trifluoromethylbenzyl)-benzamide,2,6-dichloro-N-[[4-(trifluoromethyl)-phenyl]-methyl]-benzamide,2-(2,3,3-triiodo-2-propenyl)-2H-tetrazole,2-[(1-methylethyl)-sulphonyl]-5-(trichloromethyl)-1,3,4-thiadiazole,2-[[6-deoxy-4-O-(4-O-methyl-(3-D-glycopyranosyl)-a-D-glucopyranosyl]-amino]-4-methoxy-1H-pyrrolo[2,3-d]pyrimidine-5-carbonitrile,2-aminobutane, 2-bromo-2-(bromomethyl)-pentanedinitrile,2-chloro-N-(2,3-dihydro-1,1,3-trimethyl-1H-inden-4-yl)-3-pyridinecarboxamide,2-chloro-N-(2,6-dimethylphenyl)-N-(isothiocyanatomethyl)-acetamide,2-phenylphenol (OPP),3,4-dichloro-1-[4-(difluoromethoxy)-phenyl]-pyrrole-2,5-dione,3,5-dichloro-N-[cyano[(1-methyl-2-propynyl)-oxy]-methyl]-benzamide,3-(1,1-dimethylpropyl-1-oxo-1H-indene-2-carbonitrile,3-[2-(4-chlorophenyl)-5-ethoxy-3-isoxazolidinyl]-pyridine,4-chloro-2-cyano-N,N-dimethyl-5-(4-methylphenyl)-1H-imidazole-1-sulphonamide,4-methyl-tetrazolo[1,5-a]quinazolin-5(4H)-one,8-(1,1-dimethylethyl)-N-ethyl-N-propyl-1,4-dioxaspiro[4,5]decane-2-methanamine, 8-hydroxyquinoline sulphate,9H-xanthene-2-[(phenylamino)-carbonyl]-9-carboxylic hydrazide,bis-(1-methylethyl)-3-methyl-4-[(3-methylbenzoyl)-oxy]-2,5-thiophenedicarboxylate,cis-1-(4-chlorophenyl)-2-(1H-1,2,4-triazol-1-yl)-cycloheptanol,cis-4-[3-[4-(1,1-dimethylpropyl)-phenyl-2-methylpropyl]-2,6-dimethyl-morpholinehydrochloride, ethyl[(4-chlorophenyl)-azo]-cyanoacetate, potassiumbicarbonate, methanetetrathiol-sodium salt, methyl1-(2,3-dihydro-2,2-dimethyl-inden-1-yl)-1H-imidazole-5-carboxylate,methyl N-(2,6-dimethylphenyl)-N-(5-isoxazolylcarbonyl)-DL-alaninate,methyl N-(chloroacetyl)-N-(2,6-dimethylphenyl)-DL-alaninate,N-(2,3-dichloro-4-hydroxyphenyl)-1-methyl-cyclohexanecarboxamide,N-(2,6-dimethyl phenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-furanyl)-acetamide, N-(2,6-dimethylphenyl)-2-methoxy-N-(tetrahydro-2-oxo-3-thienyl)-acetamide,N-(2-chloro-4-nitrophenyl)-4-methyl-3-nitro-benzenesulphonamide,N-(4-cyclohexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidinamine,N-(4-hexylphenyl)-1,4,5,6-tetrahydro-2-pyrimidinamine,N-(5-chloro-2-methylphenyl)-2-methoxy-N-(2-oxo-3-oxazolidinyl)-acetamide,N-(6-methoxy)-3-pyridinyl)-cyclopropanecarboxamide,N-[2,2,2-trichloro-1-[(chloroacetyl)-amino]-ethyl]-benzamide,N-[3-chloro-4,5-bis(2-propinyloxy)-phenyl]-N′-methoxy-methanimidamide,N-formyl-N-hydroxy-DL-alanine-sodium salt, O,O-diethyl[2-(dipropylamino)-2-oxoethyl]-ethylphosphoramidothioate, O-methylS-phenyl phenylpropylphosphoramidothioate, S-methyl1,2,3-benzothiadiazole-7-carbothioate, andspiro[2H]-1-benzopyrane-2,1′(3′H)-isobenzofuran]-3′-one,N-trichloromethyl)thio-4-cyclohexane-1,2-dicarboximide,tetramethylthioperoxydicarbonic diamide, methylN-(2,6-dimethylphenyl)-N-(methoxyacetyl)-DL-alaninate,4-(2,2-difluoro-1,3-benzodioxol-4-yl)-1-H-pyrrol-3-carbonitril, or anycombination thereof.

When the polypeptides are formulated or applied in combination withcommercially available fungicides, the compositions can provide an extralayer of protection for enhancing disease prevention or spread in aplant. The combination of the polypeptides with a fungicide can protecta plant against a primary or secondary fungal infection which may occurif the plant has become compromised or weakened due to exposure toabiotic stress or disease.

The strobilurin fungicide can comprise a Strobilurin A, a Strobilurin B,a Strobilurin C, a Strobilurin D, a Strobilurin E, a Strobilurin F, aStrobilurin G, a Strobilurin H, an Azoxystrobin, a Trifloxystrobin, aKresoxim methyl, a Fluoxastrobin, Picoxystrobin, or any combinationthereof.

The strobilurin fungicide can comprise a non-naturally occurringstrobilurin fungicide such as an Azoxystrobin, a Trifloxystrobin, aKresoxim methyl, a Fluoxastrobin, or any combination thereof. Forexample, the strobilurin fungicide can comprise a Trifloxystrobin,Fluoxastrobin or Picoxystrobin. Strobilurin fungicides are used tocontrol a range of fungal diseases, including water molds, downymildews, powdery mildews, leaf spotting and blighting fungi, fruitrotters, and rusts. They are useful for treating a variety of crops,including cereals, field crops, fruits, tree nuts, vegetables,turfgrasses, and ornamentals.

The triazole fungicide can comprise prothioconazole, imidazole,imidazil, prochloraz, propiconazole, triflumizole, diniconazole,flusilazole, penconazole, hexaconazole, cyproconazole, myclobutanil,tebuconazole, difenoconazole, tetraconazole, fenbuconazole,epoxiconazole, metconazole, fluquinconazole, triticonazole, or anycombination thereof.

The bioactive priming polypeptides can be delivered in combination withstrobilurins and triazole fungicides, especially fluoxastrobin ortrifloxystrobin in combination with prothioconazole.

In addition, the fungicide can comprise azoxystrobin, carboxin,difenoconazole, fludioxonil, fluxapyroxad, ipconazole, mefenoxam,pyraclostrobin, silthiofam, sedaxane, thiram, triticonazole or anycombination thereof.

In addition to foliar applied fungicides as described herein, thebioactive priming polypeptides can be provided in combination with afungicide, an insecticide, a nematicide, a bacteriocide, and a miticideor any agrochemical which is a biological agent.

The agrochemical can include a fertilizer. The fertilizer can compriseammonium sulfate, ammonium nitrate, ammonium sulfate nitrate, ammoniumchloride, ammonium bisulfate, ammonium polysulfide, ammoniumthiosulfate, aqueous ammonia, anhydrous ammonia, ammonium polyphosphate,aluminum sulfate, calcium nitrate, calcium ammonium nitrate, calciumsulfate, calcined magnesite, calcitic limestone, calcium oxide, calciumnitrate, dolomitic limestone, hydrated lime, calcium carbonate,diammonium phosphate, monoammonium phosphate, magnesium nitrate,magnesium sulfate, potassium nitrate, potassium chloride, potassiummagnesium sulfate, potassium sulfate, sodium nitrates, magnesianlimestone, magnesia, urea, urea-formaldehydes, urea ammonium nitrate,sulfur-coated urea, polymer-coated urea, isobutylidene diurea,K₂SO4-Mg₂SO₄, kainite, sylvinite, kieserite, Epsom salts, elementalsulfur, marl, ground oyster shells, fish meal, oil cakes, fish manure,blood meal, rock phosphate, super phosphates, slag, bone meal, wood ash,manure, bat guano, peat moss, compost, green sand, cottonseed meal,feather meal, crab meal, fish emulsion, humic acid, or any combinationthereof.

The fertilizer can comprise a liquid fertilizer or a dry fertilizer.

The agrochemical can comprise a micronutrient fertilizer material, themicronutrient fertilizer material comprising boric acid, a borate, aboron frit, copper sulfate, a copper frit, a copper chelate, a sodiumtetraborate decahydrate, an iron sulfate, an iron oxide, iron ammoniumsulfate, an iron frit, an iron chelate, a manganese sulfate, a manganeseoxide, a manganese chelate, a manganese chloride, a manganese frit, asodium molybdate, molybdic acid, a zinc sulfate, a zinc oxide, a zinccarbonate, a zinc frit, zinc phosphate, a zinc chelate, or anycombination thereof.

The agrochemical can comprise an insecticide, the insecticide comprisingan organophosphate, a carbamate, a pyrethroid, an acaricide, an alkylphthalate, boric acid, a borate, a fluoride, sulfur, a haloaromaticsubstituted urea, a hydrocarbon ester, a biologically-based insecticide,or any combination thereof.

When the composition includes a biostimulant, the biostimulant cancomprise a seaweed extract, an elicitor, a polysaccharide, amonosaccharide, a protein extract, a soybean extract, a humic acid, aplant hormone, a plant growth regulator, or any combination thereof.

A variety of colorants may be employed, including organic chromophoresclassified as nitroso, nitro, azo, including monoazo, bisazo, andpolyazo, diphenylmethane, triarylmethane, xanthene, methane, acridine,thiazole, thiazine, indamine, indophenol, azine, oxazine, anthraquinone,phthalocyanine, or any combination thereof.

The composition can further comprise a carrier.

The carrier of the composition can include, but is not limited to,water, peat, wheat, bran, vermiculite, clay, pasteurized soil, calciumcarbonate, calcium bicarbonate, dolomite, gypsum, bentonite, a clay, arock phosphate, a phosphorous compound, titanium dioxide, humus, talc,alginate, activated charcoal, or a combination thereof.

The composition can be in the form of an aqueous solution, a slurry ordispersion, an emulsion, a solid such as a powder or granule, or anyother desirable form for applying the composition to a plant or plantpart.

Bioactive priming polypeptides such as the flagellin andflagellin-associated polypeptides, thionin (defensin family),harpin-like HpaG, EF-Tu or other growth promoting or altering bioactivepriming polypeptides such as PSKα and RHPP can be provided ascompositions that can either be exogenously and/or endogenously appliedto a plant or a plant part and provide enhanced plant growth,productivity and enhanced health of that plant or plant part asdescribed in more detail below.

The bioactive priming polypeptides can be added separately or incombination as a composition that are useful as applications to providea benefit to plants and/or plant parts.

In combination, the polypeptides may be formulated and delivered in apurified polypeptide form either as a genetic fusion on the samerecombinant vector, or separately using different recombinant vectors.

The bioactive priming polypeptides can also be created and delivered toa plant or plant part as polypeptides from multiple actives in a fusionprotein. Examples of this include delivery of multiple flagellinassociated polypeptides produced in series with protease cleavage sitesbetween each polypeptide as is within the skill of one of ordinary skillin the art. Such fusion proteins can include any combination of thebioactive priming polypeptides as described herein, including bioactivepriming polypeptides from different classes, such as combinations offlagellin associated polypeptides with RHPP. Bioactive primingpolypeptides can also be utilized as protein fusions to plant bindingdomains, which can direct the polypeptides to distinct locations withinthe plant where they are most desired or needed for their activities tobe beneficial.

Additionally, the polypeptides may be added to formulations provided ina synthetic compound form.

The flagellin and flagellin-associated bioactive priming polypeptides asdescribed herein can be provided individually or in combinationcontaining at least two to multiple bioactive priming polypeptides toprovide a composition that meets the specific needs of a plant over awide range of desired host responses and cropping systems.

When a composition includes the retro-inverso form of a Flg bioactivepriming polypeptide (for example, RI Bt.4Q7 Flg 22 (SEQ ID NO: 376), thepolypeptide exhibits enhanced stability and less degradation over timeproviding for more activity at the plant cell membrane surface, whichenhances the ability of the polypeptide to bind to the receptor and betaken into the plant. Retro inverso forms of such Flg-associatedbioactive priming polypeptides are used to provide enhanced stability ofthe agriculturally applied formulation whereby the Flg polypeptide(s)exhibits enhanced protection from proteolytic cleavage, whichcontributes to an overall greater activity and shelf life of thecomposition.

When the polypeptide comprises an RHPP polypeptide, the composition canfurther comprise a flagellin or flagellin associated polypeptide. TheRHPP polypeptide can comprise SEQ ID NO: 600. The amino acid sequence ofthe flagellin or flagellin-associated polypeptide can comprise any oneof SEQ ID NOs: 1-525, 532, 534, 536, 538, 540, 571-586, and 751-752, orany combination thereof. For example, the flagellin or flagellinassociated polypeptide can comprise any one of SEQ ID NO: 226, 571, and752. In some instances, the RHPP polypeptide can comprise SEQ ID NO: 600and the flagellin or flagellin associated polypeptide can comprise SEQID NO: 226.

The polypeptides can be formulated in combination with an assistancepolypeptide. The signature (SEQ ID NOs: 542-548), signal anchor sorting(SEQ ID NOs: 549-562) and secretion (SEQ ID NOs: 563-570) polypeptidescan be combined with the bioactive priming polypeptides as described fortargeting the polypeptides/peptides (Tables 1-5) to the plant cellmembrane surface for improved binding and activation of theFlg-associated receptors. This means for efficient delivery and bindingof the polypeptide to a plant provides growth promoting benefits, aswell as enhanced protection to the plant or plant part.

For example, the harpin or HpaG-like bioactive priming polypeptides asdescribed herein can be used in combination with the assistancepolypeptides as described in Tables 6-8), signature polypeptides (SEQ IDNO: 542-548), signal anchor sorting (SEQ ID NO: 549-562) and/orsecretion (SEQ ID NO: 563-570) polypeptides. These assistancepolypeptides used in combination with the HpaG-like bioactive primingpolypeptides are useful to target and deliver the harpin-like bioactivepriming polypeptides to the plant cell membrane surface enhancing thecontact with the plant cell membrane and provide a conduit facilitatingefficient contact and entry of harpin-like (HpaG-like) into the plant orto the plant cell milieu (apoplast).

One or more of the EF-Tu polypeptides can be combined, optionally, withthe flagellin or flagellin-associated polypeptide. The amino acidsequence of the EF-Tu polypeptide or polypeptides can comprise SEQ IDNOs: 616 and/or 617. The amino acid sequence of the flagellin orflagellin-associated polypeptide can comprise any one of SEQ ID NOs:1-525, 532, 534, 536, 538, 540, 571-586, and 751-753 or any combinationthereof. For example, the amino acid sequence of the flagellin orflagellin-associated polypeptide can comprise SEQ ID NO: 571. As anotherexample, the composition can comprise an EF-Tu polypeptides comprisingSEQ ID NOs: 616 and 617, and a flagellin or flagellin associatedpolypeptide comprising SEQ ID NO: 226, 571, 572, or combinationsthereof. As another example, the EF-Tu polypeptide or polypeptideshaving SEQ ID NOs 616 and/or 617 can be combined with a flagellin orflagellin associated polypeptide having SEQ ID NO: 226. Alternatively,the composition can comprise one or more EF-Tu polypeptides alone (e.g.,comprising SEQ ID NOs 616 and/or 617). The EF-Tu polypeptides (e.g., SEQID Nos 616 and 617) can be further modified via N-terminal acetylation.

Additionally, the EF-Tu polypeptide or the EF-Tu polypeptide and theflagellin or flagellin-associated polypeptide can be combined with theharpin or harpin-like polypeptide. For example, the amino acid sequenceof the harpin or harpin-like polypeptide can comprise SEQ ID NO: 587.

The compositon can comprise any one of the following combinations: (a)the flagellin or flagellin-associated polypeptides and the amino acidsequences of the flagellin or flagellin-associated polypeptides compriseSEQ ID NOs: 571, 295, 300, 293, and 580; or 295, 300, 293, and 580; or571, 295, 293, and 580; or 571, 300, 293, and 580; or 571, 293 and 580;or 571, 295, 293; or (b) the flagellin or flagellin-associatedpolypeptide and the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises SEQ ID NO: 226 andcellobiose, cellulose, chitin, chitosan or any combination thereof; or(c) the flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226 and the harpin or harpin-like polypeptide and the aminoacid sequence of the harpin or harpin-like polypeptide comprises SEQ IDNO: 591; or (d) the harpin or harpin-like polypeptide and the amino acidsequence of the harpin or harpin-like polypeptide comprises SEQ ID NO:587 and the PSK polypeptide and the amino acid sequence of the PSKpolypeptide comprises SEQ ID NO: 598; or (e) the flagellin orflagellin-associated polypeptide and the amino acid sequence of theflagellin or flagellin-associated polypeptide comprises SEQ ID NO: 226,752, or 571 or any combination thereof and the EF-Tu polypeptides andthe amino acid sequences of the EF-Tu polypeptides comprise SEQ ID NOs:616 and 617; or (f) the flagellin or flagellin-associated polypeptideand the amino acid sequence of the flagellin or flagellin-associatedpolypeptide comprises SEQ ID NO: 226, 540, 752, or 571 or anycombination thereof; or (g) the RHPP polypeptide and the amino acidsequence of the RHPP polypeptide comprises SEQ ID NO: 600; or (h) theflagellin or flagellin-associated polypeptide and the amino acidsequences of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226, 540, 226, 752, or 571 or any combination thereof and theRHPP polypeptide and the amino acid sequence of the RHPP polypeptidecomprises SEQ ID NO: 600; or (i) the flagellin or flagellin-associatedpolypeptide and the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises SEQ ID NO: 226 and the RHPPpolypeptide and the amino acid sequence of the RHPP polypeptidecomprises SEQ ID NO: 600.

IV. Applications

The agricultural composition and methods described herein can be usedwith any species of plant and/or the seeds thereof. The compositions andmethods are typically used with seeds that are agronomically important.

The seed can be a transgenic seed from which a transgenic plant can growthat incorporates a transgenic event that confers, for example,tolerance to a particular herbicide or combination of herbicides,increased disease resistance, enhanced tolerance to insects, drought,stress and/or enhanced yield.

The seed can comprise a breeding trait, including for example, a diseasetolerant breeding trait.

In some instances, the seed includes at least one transgenic trait andat least one breeding trait.

The bioactive priming polypeptide compositions and methods for applyingthe polypeptides can be used for the treatment of any suitable seedtype, including, but not limited to, row crops and vegetables. Forexample, one or more plants or plant parts or the seeds of one or moreplants can comprise abaca (manila hemp) (Musa textilis), alfalfa forfodder (Medicago sativa), alfalfa for seed (Medicago sativa), almond(Prunus dulcis), anise seeds (Pimpinella anisum), apple (Malussylvestris), apricot (Prunus armeniaca), areca (betel nut) (Arecacatechu), arracha (Arracacia xanthorrhiza), arrowroot (Marantaarundinacea), artichoke (Cynara scolymus), asparagus (Asparagusofficinalis), avocado (Persea americana), bajra (pearl millet)(Pennisetum americanum), bambara groundnut (Vigna subterranea), banana(Musa paradisiaca), barley (Hordeum vulgare), beans, dry, edible, forgrains (Phaseolus vulgaris), beans, harvested green (Phaseolus and Vignaspp.), beet, fodder (mangel) (Beta vulgaris), beet, red (Beta vulgaris),beet, sugar (Beta vulgaris), beet, sugar for fodder (Beta vulgaris),beet, sugar for seeds (Beta vulgaris), bergamot (Citrus bergamia), betelnut (Areca catechu), black pepper (Piper nigrum), black wattle (Acaciamearnsii), blackberries of various species (Rubus spp.), blueberry(Vaccinium spp.), Brazil nut (Bertholletia excelsa), breadfruit(Artocarpus altilis), broad bean, dry (Vicia faba), broad bean,harvested green (Vicia faba), broccoli (Brassica oleracea var.botrytis), broom millet (Sorghum bicolor), broom sorghum (Sorghumbicolor), Brussels sprouts (Brassica oleracea var. gemmifera), buckwheat(Fagopyrum esculentum), cabbage, red, white, Savoy (Brassica oleraceavar. capitata), cabbage, Chinese (Brassica chinensis), cabbage, forfodder (Brassica spp.), cacao (cocoa) (Theobroma cacao), cantaloupe(Cucumis melo), caraway seeds (Carum carvi), cardamom (Elettariacardamomum), cardoon (Cynara cardunculus), carob (Ceratonia siliqua),carrot, edible (Daucus carota spp. sativa), carrot, for fodder (Daucuscarota sativa), cashew nuts (Anacardium occidentale), cassava (manioc)(Manihot esculenta), castor bean (Ricinus communis), cauliflower(Brassica oleracea var. botrytis), celeriac (Apium graveolens var.rapaceum), celery (Apium graveolens), chayote (Sechium edule), cherry,all varieties (Prunus spp.), chestnut (Castanea sativa), chickpea (grampea) (Cicer arietinum), chicory (Cichorium intybus), chicory for greens(Cichorium intybus), chili, dry (all varieties) (Capsicum spp.(annuum)), chili, fresh (all varieties) (Capsicum spp. (annuum)),cinnamon (Cinnamomum verum), citron (Citrus medica), citronella(Cymbopogon citrates; Cymbopogon nardus), clementine (Citrusreticulata), clove (Eugenia aromatica; Syzygium aromaticum), clover forfodder (all varieties) (Trifolium spp.), clover for seed (all varieties)(Trifolium spp.), cocoa (cacao) (Theobroma cacao), coconut (Cocosnucifera), cocoyam (Colocasia esculenta), coffee (Coffea spp.), colanut, all varieties (Cola acuminata), colza (rapeseed) (Brassica napus),corn (maize), for cereals (Zea mays), corn (maize), for silage (Zeamays), corn (maize), for vegetable (Zea mays), corn for salad(Valerianella locusta), cotton, all varieties (Gossypium spp.),cottonseed, all varieties (Gossypium spp.), cowpea, for grain (Vignaunguiculata), cowpea, harvested green (Vigna unguiculata), cranberry(Vaccinium spp.), cress (Lepidium sativum), cucumber (Cucumis sativus),currants, all varieties (Ribes spp.), custard apple (Annona reticulate),dasheen (Colocasia esculenta), dates (Phoenix dactylifera), drumsticktree (Moringa oleifera), durra (sorghum) (Sorghum bicolour), durum wheat(Triticum durum), earth pea (Vigna subterranea), edo (eddoe) (Xanthosomaspp.; Colocasia spp.), eggplant (Solanum melongena), endive (Cichoriumendivia), fennel (Foeniculum vulgare), fenugreek (Trigonellafoenum-graecum), fig (Ficus carica), filbert (hazelnut) (Corylusavellana), fique (Furcraea macrophylla), flax for fiber (Linumusitatissimum), flax for oil seed (linseed) (Linum usitatissimum),formio (New Zealand flax) (Phormium tenax), garlic, dry (Alliumsativum), garlic, green (Allium sativum), geranium (Pelargonium spp.;Geranium spp.), ginger (Zingiber officinale), gooseberry, all varieties(Ribes spp.), gourd (Lagenaria spp; Cucurbita spp.), gram pea (chickpea)(Cicer arietinum), grape (Vitis vinifera), grapefruit (Citrus paradisi),grapes for raisins (Vitis vinifera), grapes for table use (Vitisvinifera), grapes for wine (Vitis vinifera), grass esparto (Lygeumspartum), grass, orchard (Dactylis glomerata), grass, Sudan (Sorghumbicolor var. sudanense), groundnut (peanut) (Arachis hypogaea), guava(Psidium guajava), guinea corn (sorghum) (Sorghum bicolor), hazelnut(filbert) (Corylus avellana), hemp fiber (Cannabis sativa spp. indica),hemp, manila (abaca) (Musa textilis), hemp, sun (Crotalaria juncea),hempseed (marijuana) (Cannabis sativa), henequen (Agave fourcroydes),henna (Lawsonia inermis), hop (Humulus lupulus), horse bean (Viciafaba), horseradish (Armoracia rusticana), hybrid maize (Zea mays),indigo (Indigofera tinctoria), jasmine (Jasminum spp.), Jerusalemartichoke (Helianthus tuberosus), jowar (sorghum) (Sorghum bicolor),jute (Corchorus spp.), kale (Brassica oleracea var. acephala), kapok(Ceiba pentandra), kenaf (Hibiscus cannabinus), kohlrabi (Brassicaoleracea var. gongylodes), lavender (Lavandula spp.), leek (Alliumampeloprasum; Allium porrum), lemon (Citrus limon), lemongrass(Cymbopogon citratus), lentil (Lens culinaris), lespedeza, all varieties(Lespedeza spp.), lettuce (Lactuca sativa var. capitata), lime, sour(Citrus aurantifolia), lime, sweet (Citrus limetta), linseed (flax foroil seed) (Linum usitatissimum), licorice (Glycyrrhiza glabra), litchi(Litchi chinensis), Ioquat (Eriobotrya japonica), lupine, all varieties(Lupinus spp.), Macadamia (Queensland nut) (Macadamia spp. ternifolia),mace (Myristica fragrans), maguey (Agave atrovirens), maize (corn) (Zeamays), maize (corn) for silage (Zea mays), maize (hybrid) (Zea mays),maize, ordinary (Zea mays), mandarin (Citrus reticulata), mangel (fodderbeet) (Beta vulgaris), mango (Mangifera indica), manioc (cassava)(Manihot esculenta), maslin (mixed cereals) (mixture of Triticum spp.and Secale cereale), medlar (Mespilus germanica), melon, exceptwatermelon (Cucumis melo), millet broom (Sorghum bicolor), millet, bajra(Pennisetum americanum), millet, bulrush (Pennisetum americanum),millet, finger (Eleusine coracana), millet, foxtail (Setaria italica),millet, Japanese (Echinochloa esculenta), millet, pearl (bajra, bulrush)(Pennisetum americanum), millet, proso (Panicum miliaceum), mint, allvarieties (Mentha spp.), mulberry for fruit, all varieties (Morus spp.),mulberry for silkworms (Morus alba), mushrooms (Agaricus spp.; Pleurotusspp.; Volvariella), mustard (Brassica nigra; Sinapis alba), nectarine(Prunus persica var. nectarina), New Zealand flax (formio) (Phormiumtenax), Niger seed (Guizotia abyssinica), nutmeg (Myristica fragrans),oats, for fodder (Avena spp.), oil palm (Elaeis guineensis), okra(Abelmoschus esculentus), olive (Olea europaea), onion seed (Alliumcepa), onion, dry (Allium cepa), onion, green (Allium cepa), opium(Papaver somniferum), orange (Citrus sinensis), orange, bitter (Citrusaurantium), ornamental plants (various), palm palmyra (Borassusflabellifer), palm, kernel oil (Elaeis guineensis), palm, oil (Elaeisguineensis), palm, sago (Metroxylon sagu), papaya (pawpaw) (Caricapapaya), parsnip (Pastinaca sativa), pea, edible dry, for grain (Pisumsativum), pea, harvested green (Pisum sativum), peach (Prunus persica),peanut (groundnut) (Arachis hypogaea), pear (Pyrus communis), pecan nut(Carya illinoensis), pepper, black (Piper nigrum), pepper, dry (Capsicumspp.), persimmon (Diospyros kaki; Diospyros virginiana), pigeon pea(Cajanus cajan), pineapple (Ananas comosus), pistachio nut (Pistaciavera), plantain (Musa sapientum), plum (Prunus domestica), pomegranate(Punica granatum), pomelo (Citrus grandis), poppy seed (Papaversomniferum), potato (Solamum tuberosum), palm, kernel oil (Elaeisguineensis), potato, sweet (Ipomoea batatas), prune (Prunus domestica),pumpkin, edible (Cucurbita spp.), pumpkin, for fodder (Cucurbita spp.),pyrethum (Chrysanthemum cinerariaefolium), quebracho (Aspidospermaspp.), Queensland nut (Macadamia spp. ternifolia), quince (Cydoniaoblonga), quinine (Cinchona spp.), quinoa (Chenopodium quinoa), ramie(Boehmeria nivea), rapeseed (colza) (Brassica napus), raspberry, allvarieties (Rubus spp.), red beet (Beta vulgaris), redtop (Agrostisspp.), rhea (Boehmeria nivea), rhubarb (Rheum spp.), rice (Oryza sativa;Oryza glaberrima), rose (Rose spp.), rubber (Hevea brasiliensis),rutabaga (swede) (Brassica napus var. napobrassica), rye (Secalecereale), ryegrass seed (Lolium spp.), safflower (Carthamus tinctorius),sainfoin (Onobrychis viciifolia), salsify (Tragopogon porrifolius),sapodilla (Achras sapota), satsuma (mandarin/tangerine) (Citrusreticulata), scorzonera (black salsify) (Scorzonera hispanica), sesame(Sesamum indicum), shea butter (nut) (Vitellaria paradoxa), sisal (Agavesisalana), sorghum (Sorghum bicolor), sorghum, broom (Sorghum bicolor),sorghum, durra (Sorghum bicolor), sorghum, guinea corn (Sorghumbicolor), sorghum, jowar (Sorghum bicolor), sorghum, sweet (Sorghumbicolor), soybean (Glycine max), soybean hay (Glycine max), spelt wheat(Triticum spelta), spinach (Spinacia oleracea), squash (Cucurbita spp.),strawberry (Fragaria spp.), sugar beet (Beta vulgaris), sugar beet forfodder (Beta vulgaris), sugar beet for seed (Beta vulgaris), sugarcanefor fodder (Saccharum officinarum), sugarcane for sugar or alcohol(Saccharum officinarum), sugarcane for thatching (Saccharumofficinarum), sunflower for fodder (Helianthus annuus), sunflower foroil seed (Helianthus annuus), sunhemp (Crotalaria juncea), swede(Brassica napus var. napobrassica), swede for fodder (Brassica napusvar. napobrassica), sweet corn (Zea mays), sweet lime (Citrus limetta),sweet pepper (Capsicum annuum), sweet potato (Lopmoea batatas), sweetsorghum (Sorghum bicolor), tangerine (Citrus reticulata), tannic(Xanthosoma sagittifolium), tapioca (cassava) (Manihot esculenta), taro(Colocasia esculenta), tea (Camellia sinensis), teff (Eragrostisabyssinica), timothy (Phleum pratense), tobacco (Nicotiana tabacum),tomato (Lycopersicon esculentum), trefoil (Lotus spp.), triticale, forfodder (hybrid of Triticum aestivum and Secale cereale), tung tree(Aleurites spp.; Fordii), turnip, edible (Brassica rapa), turnip, forfodder (Brassica rapa), urena (Congo jute) (Urena lobata), vanilla(Vanilla planifolia), vetch, for grain (Vicia sativa), walnut (Juglansspp., especially Juglans regia), watermelon (Citrullus lanatus), wheat(Triticum aestivum), yam (Dioscorea spp.), or yerba mate (Ilexparaguariensis).

The compositions and methods disclosed herein can also be applied toturf grass, ornamental grass, flowers, ornamentals, trees, and shrubs.

The compositions comprising the bioactive priming polypeptides are alsosuitable for use in the nursery, lawn and garden, floriculture or thecut flower industry and provide benefits for enhanced plantproductivity, protection health, vigor and longevity. For example, theycan be applied to perennials, annuals, forced bulbs, or pseudo bulbs,herbs, groundcovers, trees, shrubs, ornamentals (e.g., orchids, etc.),tropicals, and nursery stock.

The compositions comprising the bioactive priming polypeptides aresuitable for treating plants, plant parts and plant propagationmaterial(s), for example, any plant or plant part, such as seeds, roots,stems, floral organs, root stocks, scions, bulb, pseudobulbs, rhizomes,tubers, etc.

The bioactive priming polypeptides can be applied as seed treatments totreat for a number of pests, diseases, nutrient deficiencies whileenhancing plant growth and productivity.

Seed coating or dressing compositions can be, for example, a liquidcarrier composition, a slurry composition, or a powder compositionapplied with conventional additives that are provided to make the seedtreatment have sticky qualities to stick to and coat the seeds. Suitableadditives for a seed composition comprise: talcs, graphites, gums,stabilizing polymers, coating polymers, finishing polymers, slip agentsfor seed flow and plantability, cosmetic agents and cellulosic materialssuch as carboxymethyl cellulose and the like. The bioactive primingpolypeptide seed treatments can further comprise colorant agents andother such additives.

The bioactive priming polypeptides can be applied individually as seedtreatments or in combination with other additives such as fungicides,insecticides, inoculants, plant growth regulators, plant growthpromoting microbes, fertilizers and fertilizer enhancers, seednutrients, biological control agents, herbicidal antidotes and seedlingdisease treatments and with other conventional seed treatments.

The seed treatment composition as described herein can be applied toseeds in a suitable carrier such as water or a powder that is notharmful to the seeds or the environment. The seeds are then planted inconventional fashion.

Preferred seed treatments such as Bt.4Q7Flg22 (SEQ ID NO: 226 or SEQ IDNO: 571), Ec.Flg22 (SEQ ID NO: 526) and Gm.RHPP (SEQ ID NO: 600) areuseful to enhance seedling development, decrease the time forgermination, increase the number of seeds that germinate, and enhanceseedling survivability. In addition, the seed treatment compositionsenhance seed protection from microbial-based diseases which are known tocontact the seed or the soil surrounding the seed and spread duringearly seedling establishment.

The seed treatment composition can comprise a polypeptide as describedherein and a fungicide, an insecticide, a nematocide, a biologicalcontrol agent, a biostimulant, a microbe, or any combination thereof.

The seed treatment composition can comprise a polypeptide as describedherein and clothianidin, Bacillus firmus, metalaxyl, or any combinationthereof.

The seed treatment composition can comprise a polypeptide as describedherein, clothianidin and fluopyram.

The seed treatment can comprise a polypeptide as described herein,metalaxyl and fluopyram.

The bioactive priming polypeptides can be applied directly to the seedas a solution or in combination with other commercially availableadditives. Solutions containing the water-soluble polypeptide can besprayed or otherwise applied to the seed as a seed slurry or a seedsoak. Solids or dry materials containing soluble bioactive primingpolypeptides are also useful to promote effective seedling germination,growth and protection during early seedling establishment.

The bioactive priming polypeptides can be formulated with a solubilizingcarrier such as water, buffer (e.g., citrate or phosphate buffer) andother treating agents (i.e., alcohol, other solvents) or anysolubilizing agent. In addition, small amounts of drying agentenhancers, such as lower alcohols, etc. can be utilized in thecomposition. Surfactants, emulsifiers and preservatives can also beadded at small (0.5% v/v or less) levels in order to enhance thestability of the seed coating product.

Seed treatments containing the bioactive priming polypeptides can beapplied using any commercially available seed treatment machinery or canalso be applied using any acceptable non-commercial method(s) such asthe use of syringes or any other seed treatment device. General seedtreatments coating procedures using bioactive priming polypeptides canbe performed using a Wintersteiger HEGE 11 (Wintersteiger AG, Austria,Germany) and applied to the seed of major crops, namely corn, soybean,wheat, rice and various vegetables. The capacity of this seed treatmentmachinery can accommodate a large number of different seed types, sizesand amounts of seed (20-3000 grams). The seed is loaded into bowls ofthe seed treater machinery. The bowl selection depends on the treatmentseed amount required and the size of the bowl selected: large 14.5 Lbowl (500-3000 g seed per coating); medium 7 L bowl (80-800 g seed percoating); and small 1 L bowl (20-100 g seed per coating). Other largerseed treatment systems are also available.

The seed is distributed toward the radial peripheries of the rotatablebowls via an application of centrifugal force with the centrifugalcoating device. The spinning disc located at the bottom of the bowldistributes the seed treatment evenly over the seed. At this point, thespin cycle is started which causes the seeds to revolve around the bowlcenter in a circle to evenly coat the seeds. The process of seedtreatment coating is initiated after the seed is evenly dispersed aroundthe spreader. Seed treatment sample material (such as a powdered,semi-liquid, liquid or a slurry) can be applied onto the rotatable diskas the disks are spinning within the rotatable bowls used to distributethe seed treatment evenly to provide a uniform coat and dress thesurface of the seed.

A constant air flow delivered using compressed air (2-6 bars) can beprovided during seed coating to assist with uniformly coating the seedsin the bowl. The amount of time for the coating of the seed depends onthe amount of the seed, the viscosity of the seed treatment and the typeof the seed used in the treatment. A seed treatment calculator is usedto adjust for all volumes, for most major and commercially grown cropsand the type of seed treatment being applied.

The seeds can be coated using a variety of methods including, but notlimited to, pouring or pumping, drizzling or spraying an aqueoussolution containing the bioactive priming polypeptides on or over aseed, spraying or applying onto a layer of seeds either with the use orwithout the use of a conveyor system. Suitable mixing devices includetumblers, mixing basins or drums, or other fluid applicating devicesthat include basins or drums used to contain the seed while coating.

After the seed has been treated and dried, the seeds are distributedinto a larger storage container(s). Seeds are either air dried or driedwith a continuous air stream that passes over the seeds. Seeds are thentransferred into a separate container or bag for shipment, transfer orstorage.

The bioactive priming polypeptides can further be provided for deliveryto a plant surface or plant plasma membrane as a foliar spray or a seedtreatment to an area surrounding a plant or a plant part.

The bioactive priming polypeptide formulation(s) can also be provided asa seed treatment application or on a matrix such as immobilized orimpregnated on a particle, or a granule such as used in a broadcasttreatment.

The bioactive priming polypeptides as described herein can be applied toplants and plant parts using an exogenous application as a spray, soiltreatment, in furrow, seed treatment, dip or wash or as an endogenousapplication as an injection, inoculation, irrigation, infiltration, etc.

The polypeptides can be applied directly to a plant or to the areasurrounding a plant or plant part.

They can also be provided on a matrix material which is then provided toa plant or plant part.

The compositions containing the flagellin-associated bioactive primingpolypeptides can also be provided for direct delivery into a plant,plant tissues or a plant cell by various delivery methods, for example,injection, inoculation or infiltration (for example, infiltration intothe stomata on the leaf). These polypeptides can also be provided in amanner where they can move systemically through a plant and influencesignaling cascades in the plant that subsequently produce beneficial andproductive outcomes to the plant or plant part.

Retro-inverso Flg bioactive priming polypeptides as described in Table 4or Table 5 can be applied individually or in combination with any otherflagellin, flagellin-associated or other bioactive priming polypeptidesequences as described herein. Combinations of such RI flagellin andflagellin-associated bioactive priming polypeptides are useful as plantprotectants as well as plant growth promoting enhancers.

The signature (SEQ ID NO: 542-548; Table 6), signal anchor sorting (SEQID NO: 549-562, Table 7) and secretion assistance polypeptides (SEQ IDNOs 563-570; Table 8) can be used in combination with any of theflagellin coding (Table 1), N and/or C-terminal conserved sequences fromBacillus-derived flagellins (Table 2), flagellin-associatedpolypeptides: Flg22 and FlgII-28 (Table 3), the retro inverso forms ofFlg22 and FlgII-28 (Table 4) or any of the other Flgs (Table 5) asdescribed herein.

For example, any of the Flg-associated bioactive priming polypeptides orcombinations thereof can be provided in individual formulations andapplied either simultaneously, sequentially in separate formulations orprovided as fusion protein(s) that contain the assistance sequences asdescribed in Tables 6-8 and applied directly or separately to a plant orplant part.

Harpin-like polypeptides or RHPP polypeptides can provide functionalbenefits when applied both exogenously, for example as a foliar spray tothe plant surface, or provided apoplastically (to the space outside ofthe plant cell membrane) or endogenously (inside a plant cell/plant cellmembrane). RHPP polypeptides can also provide functional benefits whenapplied as a seed treatment.

Foliar or in furrow applications of harpin-like, HpaG-like polypeptidesare useful to enhance growth, increase biomass, and greenness orchlorophyll production of a plant.

The PSKα bioactive priming polypeptide(s) can be provided for deliveryto a plant surface/plant plasma membrane as a foliar spray or, a seedtreatment to an area surrounding a plant, plant part or a plant cell.

The compositions containing the PSKα bioactive priming polypeptides canalso be provided for delivery into a plant, plant tissues or a plantcell by various delivery methods, for example, injection, inoculation orinfiltration (for example, added directly or prerequisitely to cellculture).

V. Methods of Use

Methods are provided for increasing growth, yield, health, longevity,productivity, and/or vigor of a plant or a plant part and/or decreasingabiotic stress in the plant or the plant part and/or protecting theplant or the plant part from disease, insects and/or nematodes, and/orincreasing the innate immune response of the plant or the plant partand/or changing plant architecture. The method can comprise applying thepolypeptide or the composition as described herein to a plant, a plantpart, or a plant growth medium or a rhizosphere in an area surroundingthe plant or the plant part to increase growth, yield, health,longevity, productivity, and/or vigor of the plant or the plant partand/or decrease abiotic stress in the plant or the plant part and/orprotect the plant or the plant part from disease, insects and/ornematodes, and/or increase the innate immune response of the plant orthe plant part and/or change the plant architecture.

Alternatively, the method can comprise applying the polypeptide or thecomposition as described herein to a plant growth medium to increasegrowth, yield, health, longevity, productivity, and/or vigor of a plantor a plant part to be grown in the plant growth medium and/or decreaseabiotic stress in the plant or the plant part to be grown in the plantgrowth medium and/or protect the plant or the plant part to be grown inthe plant growth medium from disease, insects and/or nematodes, and/orincrease the innate immune response and/or change plant architecture ofthe plant or the plant part to be grown in the plant growth medium.

Another method comprises applying the recombinant microorganism asdescribed herein to a plant, a plant part, or a plant growth medium or arhizosphere in an area surrounding the plant or the plant part toincrease growth, yield, health, longevity, productivity, and/or vigor ofthe plant or the plant part and/or decrease abiotic stress in the plantor the plant part and/or protect the plant or the plant part fromdisease, insects and/or nematodes, and/or increase the innate immuneresponse of the plant or the plant part and/or change the plantarchitecture. The recombinant microorganism expresses the polypeptideand expression of the polypeptide is increased as compared to theexpression level the polypeptide in a wild-type microorganism of thesame kind under the same conditions.

Methods using the bioactive priming polypeptides are also provided toincrease the overall plant productivity in a field, orchard, plantingbed, nursery, timberland, farm, lawn, garden, garden center or acreage.Applications and methods using the bioactive priming polypeptides arealso useful for increasing plant growth, health and productivity indiverse crops (monocots and dicots), for example, corn, wheat, rice,sugarcane, soybean, sorghum, potatoes and a variety of vegetables.

A “bioactive polypeptide priming” approach is also provided by directapplication of the polypeptides, which can be applied either exogenouslyto a plant cell surface or endogenously to the interior of a plantand/or a plant cell. The polypeptides are provided for delivery to theplant surface or plasma cell membrane or to the interior of a plant,plant tissue or cell and are useful for regulating developmentalprocesses that result in enhanced growth phenotypes such as increases inoverall biomass, vegetative growth, seed fill, seed size, and number ofseed that contribute to increases in the total yield of crop plants.

Application of the retro-inverso Flg polypeptides provided inagricultural formulations can result in enhanced plant protection fromdiseases and abiotic stresses while synergistically enhancing growth,productivity and yield while maintaining increased plant health withenhanced plant performance for longer periods of time.

Selection of the native L (Table 3) or the retro-inverso D (Table 4)forms of the Flg-associated polypeptides can depend on the environment,the plant/crop, or the combination of plant/crop and environment. Inaddition, the timing of the treatment application (for example, a foliarspray application) during the growing season are all relevantconsiderations. The retro inverso Flg bioactive priming polypeptideshave enhanced binding affinity to cell surface membranes. Due to thesefeatures, the RI forms of the Flg bioactive priming polypeptides can beused to improve abiotic stress tolerance in a plant or plant part.

Additionally, the retro inverso forms of RI Ec.Flg22 and RI Bt.4Q7Flg22can be useful to stimulate the closure of stomata under conditions ofdrought and heat stress and improve yields under those conditions.Control of stomatal closure using Flg-associated bioactive primingpolypeptide applied to a plant during periods of environmental stresscan assist in the regulation of water loss and stabilize turgor pressurein a plant when environmental conditions are unfavorable.

In the methods, the polypeptide or the composition can comprise: theFlg22 polypeptide and an amino acid sequence of the Flg22 polypeptidecomprising any one of SEQ ID NOs: 226-300 and 571-573; the retro inversoFlg22 polypeptide and an amino acid sequence of the retro inverso Flg22polypeptide comprising any one of SEQ ID NO: 376-450; or any combinationthereof to protect the plant or the plant part from disease and/orincrease the innate immune response of the plant or the plant part.

In the methods, the polypeptide or the composition can comprise: theFlgII-28 polypeptide and an amino acid sequence of the FlgII-28polypeptide comprising any one of SEQ ID NOs: 301-375; the retro inversoFlgII-28 polypeptide and an amino acid sequence of the retro inversoFlgII-28 polypeptide comprising any one of SEQ ID NO: 451-525; or anycombination thereof to protect the plant or the plant part from diseaseand/or increase the innate immune response of the plant or the plantpart.

In the methods, the polypeptide or the composition can comprise theFlgII-28 polypeptide and an amino acid sequence of the Flg22 polypeptidecan comprise any one of SEQ ID NO: 226, 571, or 752 and/or EF-Tupolypeptides, the amino acid sequence of the EF-Tu polypeptidescomprising SEQ ID NOs: 616 and 617, to protect the plant or the plantpart from disease and/or increase the innate immunity of the plant orplant part. In the methods, the amino acid sequence of the flagellin orflagellin-associated polypeptide can comprise any one of SEQ ID NOs:226, 289, 290, 291, 293, 294, 295, 300, 437, 532, 534, 536, 538, 540,571-586, and 751-766 or any combination thereof to protect the plant orthe plant part from disease, insects or nematodes. These arepolypeptides with mutant sequences exhibiting increased activity toreactive oxygen species. For example, the amino acid sequence of theflagellin or flagellin-associated polypeptide can comprise any one ofSEQ ID NOs: 226, 293, 295, 300, 540, 571 574, 751 and 752 or anycombination thereof.

The disease can comprise Asian citrus greening, Huanglonging (HLB)disease, Asian soybean rust, Sclerotinia stem rot (or white mold),Pseudomonas leaf spot, or Cercospora leaf blight.

In the methods, the polypeptide or the composition can comprise theFlg22 polypeptide and an amino acid sequence of the Flg22 polypeptidecomprising any one of SEQ ID NOs: 226-300 and 571-573 or any combinationthereof.

In the methods, the polypeptide or the composition can comprise theFlgII-28 polypeptide and an amino acid sequence of the FlgII-28polypeptide comprising any one of SEQ ID NOs: 301-375 or 751 or anycombination thereof.

In the methods, the polypeptide or the composition can comprise theFlg22 polypeptide and the FlgII-28 polypeptide, an amino acid sequenceof the Flg22 polypeptide comprising any one of SEQ ID NOs: 226-300 and571-573 or any combination thereof and an amino acid sequence of theFlgII-28 polypeptide comprising any one of SEQ ID NOs: 301-375 or 751 orany combination thereof. The polypeptide or the composition can furthercomprise the retro inverso Flg22 polypeptide, the retro inverso FlgII-28polypeptide or a combination thereof, an amino acid sequence of theretro inverso Flg22 polypeptide comprising any one of SEQ ID NO: 376-450or any combination thereof and an amino acid sequence of the retroinverso FlgII-28 polypeptide comprising any one of SEQ ID NO: 451-525 orany combination thereof.

In the methods, the polypeptide or the composition can comprise the RHPPpolypeptide and/or the RI RHPP polypeptide to increase the yield, thegrowth and/or the productivity of the plant or plant part and/or changethe plant architecture.

When the method includes a polypeptide or composition comprising theRHPP polypeptide and/or the RI RHPP polypeptide, the growth can compriseroot growth, root length, root biomass, nodulation, total biomass, aboveground biomass, or any combination thereof. When the polypeptide orcomposition comprises the RHPP polypeptide, the amino acid sequence ofthe RHPP polypeptide can comprise SEQ ID NO: 600.

When the method includes a polypeptide or composition comprising theRHPP polypeptide and/or the RI RHPP polypeptide, the plant can comprisesoybean, the growth can comprise overall root length, root biomass,nodulation, nodules per plant, total biomass, above ground biomass, orany combination thereof, and the productivity can comprise number oftotal pods or pods per node.

The plant architecture can comprise beneficial outcomes to the plant orplant part. For example, the beneficial outcomes can include increasedplanting density capability for a field of the plants.

In the methods, the polypeptide or the composition can comprise theharpin-like polypeptide or the RHPP polypeptide to protect the plant orthe plant part from disease, insects and/or nematodes, and/or increasethe innate immune response of the plant or the plant part.

In the methods, the polypeptide or the composition can comprise the PSKpolypeptide to increase yield of the plant or the plant part inenvironments prone to heat and drought.

The polypeptide, the composition, or the recombinant microorganism canbe applied just prior to floral formation or at the pre-flowering stage.

In the methods, the polypeptide or the composition can comprise the PSKpolypeptide, the RHPP, the harpin or harpin-like polypeptide, or acombination thereof to increase growth of the plant or the plant part.

The growth can comprise root and floral apical meristems, floral organproduction, fruit development, fruit production, number of floralorgans, size of floral organs, or a combination thereof.

In the methods, the polypeptide or the composition can comprise the PSKpolypeptide and the harpin or harpin-like polypeptide to increase growthand productivity of the plant or the plant part in an environment proneto both stress and non-stress conditions for plant growth.

In the methods, the polypeptide or the composition can comprise thethionin or thionin-like polypeptide.

The thionin or thionin-like polypeptide can be fused to a phloemtargeting sequence to form a fused polypeptide, the amino acid sequenceof the phloem targeting sequence comprising any one of SEQ ID NOs:641-649, or any combination thereof, for delivering the fusedpolypeptide to vascular tissue or cells and/or phloem orphloem-associated tissue or cells in the plant or plant part.

In the methods, protecting the plant or the plant part from disease cancomprise prophylactic treatment, treatment, prevention and decreaseddisease progression on or in the plant or plant part.

The disease can comprise Asian citrus greening disease (HLB), Citruscanker disease, Cercospora leaf blight or a bacteria causing disease.

The bacteria causing disease can comprise bacterial leaf blight,bacterial leaf streak, bacterial stalk rot, bacterial leaf spot,bacterial leaf scorch, bacterial top rot, bacterial stripe, chocolatespot, Goss's bacterial wilt and blight, Holcus spot, purple leaf sheath,seed rot, seedling blight, Stewart's disease (bacterial wilt), cornstunt, Fire Blight, Pierce's disease, citrus variegated chlorosis,citrus canker, Pseudomonas syringae serovars, or a combination thereof.

In the methods, the polypeptide or the composition further can comprisethe flagellin or flagellin-like polypeptide, and an amino acid sequenceof the flagellin or flagellin-like polypeptide comprising any one of SEQID NOs: 226-525 and 571-573 or any combination thereof.

In the methods, the polypeptide, the composition, or the recombinantmicroorganism can be applied exogenously to the plant, the plant part,or the plant growth medium.

In the methods, the polypeptide, the composition, or the recombinantmicroorganism can be applied endogenously to the plant or the plantpart.

The plant part can include a plant cell, a leaf, a branch, a stem, aflower, a foliage, a floral organ, a fruit, pollen, a vegetable, atuber, a rhizome, a corm, a bulb, a pseudobulb, a pod, a root, a rootball, a root stock, a scion, or a seed.

In the methods, the polypeptide, the composition, or the recombinantmicroorganism can be applied to a surface of the plant, a foliage of theplant or a surface of a seed of the plant.

In the methods, the polypeptide, the composition, or the recombinantmicroorganism can be applied to the surface of the seed and the plant orthe plant part is grown from the seed.

In the methods, the polypeptide, the composition, or the recombinantmicroorganism can be applied as a foliar application.

The plant can be a fruit plant or a vegetable plant, and the methodprovides increased yield of fruits or vegetables.

In methods where the bioactive priming polypeptides are applied two ormore times during a growing season, the first application can occur ator before the V2 stage of development, and subsequent applications canoccur before the plant flowers. For example, the first application canoccur as a seed treatments, at/or before the VE stage of development, ator before the V1 stage of development, at or before the V2 stage ofdevelopment, at or before the V3 stage of development, at or before theV4 stage of development, at or before the V5 stage of development, at orbefore the V6 stage of development, at or before the V7 stage ofdevelopment, at or before the V8 stage of development, at or before theV9 stage of development, at or before the V10 stage of development, ator before the V11 stage of development, at or before the V12 stage ofdevelopment, at or before the V13 stage of development, at or before theV14 stage of development, at or before the V15 stage of development, ator before the VT stage of development, at or before the R1 stage ofdevelopment, at or before the R2 stage of development, at or before theR3 stage of development, at or before the R4 stage of development, at orbefore the R5 stage of development, at or before the R6 stage ofdevelopment, at or before the R7 stage of development, or at or beforethe R8 stage of development. By way of example, the first applicationcan occur at or before the germination stage, at or before the seedlingstage, at or before the tillering stage, at or before the stemelongation stage, at or before the booting stage, or at or before theheading stage. For example, where the Feekes scale is used to identifythe stage of growth of a cereal crop, the first application can occur ator before stage 1, at or before stage 2, at or before stage 3, at orbefore stage 4, at or before stage 5, at or before stage 6, at or beforestage 7, at or before stage 8, at or before stage 9, at or before stage10, at or before stage 10.1, at or before stage 10.2, at or before stage10.3, at or before stage 10.4, or at or before stage 10.5.

Abiotic Stress

Abiotic stress causes significant crop loss and can result in majorreductions in crop production and yield potential. The bioactive primingpolypeptides and compositions as described herein can be used aschemical priming agents to increase tolerance of a plant to one or moreabiotic stresses. Thus, the flagellin polypeptides, flagellin-associatedpolypeptides of Flg22 or FlgII-28 derived from Bacillus species, Flg15and Flg22 derived from E. coli and other organisms (Table 5) and theRHPP polypeptides derived from Glycine max (Tables 13 to 15) are usefulfor increasing the tolerance of a plant, group of plants, field ofplants and/or the parts of plants to abiotic stress. The polypeptidesand compositions as described herein impart abiotic stress tolerance toa plant or plant part. The abiotic stress tolerance imparted to a plantor plant part are to abiotic stresses that include, but are not limitedto: temperature stress, radiation stress, drought stress, cold stress,salt stress, osmotic stress, nutrient-deficient or high metal stress,and water stress that results from water deficit, flooding or anoxia.Chemical priming using the bioactive priming polypeptides andcompositions as described herein are applied to a plant or plant partoffering a versatile approach to protect the plant or plant part againstindividual, multiple or combined abiotic stresses.

The polypeptides and compositions as described herein are effective toprotect a plant against abiotic stressors when applied as an aboveground foliar application to a plant, a plant part, a plant root, aplant seed, a plant growth medium, or the area surrounding a plant orthe area surrounding a plant seed.For example, for trees, one or moreapplications can be applied at different growth timings of trees,including timings before, during or after flushes; before, during, orafter fruit set; or before or after fruit harvest.

The methods described herein chemically prime the plant for protectionagainst abiotic stress(es) in such a way that the plant has alreadyprepared and initiated defense mechanisms that can be activated fasterand increase tolerance to an abiotic stress or multiple stressorsoccurring simultaneously or at different times during the growingseason.

The retro inverso forms of the Flg22 polypeptides as described hereincan be applied externally as a foliar spray application (or using otherapplication methods as well, for example as a root drench) during timesof excessive heat, water, and drought stress and be used to protect aplant against drought, heat stress and/or other abiotic stresses thatcan affect stomatal aperture and oscillation that commonly occur withtranspiration loss through a plant.

In the methods, the polypeptide or the composition can comprise: theFlg22 polypeptide and an amino acid sequence of the Flg22 polypeptidecomprising any one of SEQ ID NOs: 226-300 and 571-573 or any combinationthereof; the retro inverso Flg22 polypeptide and an amino acid sequenceof the retro inverso Flg22 polypeptide comprising any one of SEQ ID NO:376-450 or any combination thereof; or any combination thereof todecrease abiotic stress in the plant or the plant part and/or protectthe plant or the plant part from disease and/or increase the innateimmune response of the plant or the plant part.

In the methods, the polypeptide or the composition can comprise: theFlgII-28 polypeptide and an amino acid sequence of the FlgII-28polypeptide comprising any one of SEQ ID NOs: 301-375 or any combinationthereof; the retro inverso FlgII-28 polypeptide and an amino acidsequence of the retro inverso FlgII-28 polypeptide comprising any one ofSEQ ID NO: 451-525 or any combination thereof; or any combinationthereof to decrease abiotic stress in the plant or the plant part and/orprotect the plant or the plant part from disease and/or increase theinnate immune response of the plant or the plant part.

In the methods, the polypeptide or the composition can comprise: theretro inverso Flg22 polypeptide and an amino acid sequence of the retroinverso Flg22 polypeptide comprising any one of SEQ ID NO: 376-450 orany combination thereof; the retro inverso FlgII-28 polypeptide and anamino acid sequence of the retro inverso FlgII-28 polypeptide comprisingany one of SEQ ID NO: 451-525 or any combination thereof; or anycombination thereof to decrease abiotic stress in the plant or the plantpart and/or protect the plant or the plant part from disease and/orincrease the innate immune response of the plant or the plant part.

In the methods, the polypeptide or the composition can comprise the RHPPpolypeptide and an amino acid sequence of the RHPP polypeptide comprisesSEQ ID NO: 600, 603, 604 or any combination thereof; the Kunitz TrypsinInhibitor (KTI) polypeptide and an amino acid sequence of the KTIpolypeptide comprises SEQ ID NO: 602; the retro-inverso RHPP polypeptideand an amino acid sequence of the RI RHPP comprises SEQ ID NO 601, 605,606 or any combination thereof; or any combination thereof to decreaseabiotic stress in the plant or the plant part and/or protect the plantor the plant part from disease and/or increase the innate immuneresponse of the plant or the plant part.

The abiotic stress can comprise heat stress, temperature stress,radiation stress, drought stress, cold stress, salt stress,nutrient-deficient stress, high metal stress, water stress, osmoticstress, or any combination thereof.

Balancing Immune Response with Plant Growth and Development

Although immune responses can provide protection of plants from pathogenattack, excessive immune responses may have negative impacts on plantgrowth. Therefore, balancing enhanced immunity or disease prevention andprotection in a plant with an increased growth promoting response is adesired combination to optimize plant health.

Bioactive priming polypeptides that are useful for enhancing immuneresponses as described herein can be combined with polypeptides thatprovide positive impacts on plant growth and productivity. Thepolypeptide combinations are specifically selected for their distinctmodes of action/regulation when applied to a plant or plant part.However, some of the bioactive priming polypeptides (Flgs, HpGa-like,PSKα, thionins) are perceived by receptor-like proteins, followed by aprocess that initiates their entry and transport in the plant whichresults in functional outcomes while others are taken into the plant byactive absorption (e.g., RHPP). For example, PSKα and the Flg-associatedpolypeptides such as Flg22, Flg25 and FlgII-28 are perceived by aleucine-rich receptor kinase located on the surface of the plasmamembrane and involve a complex signaling pathway involved in thepathogen-triggered responses leading to immunity, disease resistance ordisease prevention (Kutschmar et al. “PSKα promotes root growth inArabidopsis,” New Phytologist 181: 820-831, 2009).

The bioactive priming polypeptides as described herein such as Flg22HpaG-like polypeptides and thionins can act as elicitors and exhibitantimicrobial activity (e.g., anti-pesticide; bacterial, fungal, orviral activity). Specific combinations of polypeptides are provided, forexample, the combination of flagellin- and harpin-associated bioactivepriming polypeptides are useful for preventing and protecting plantsfrom pathogenic diseases and serve a dual utility when they are appliedtogether with those other polypeptides, for example, PSKα and RHPP, thatenhance plant growth and productivity in a plant, plant part, and/orfield of plants.

The combinations of bioactive priming polypeptides as described hereincan be applied exogenously as a foliar spray, in furrow treatment, seedtreatment, drench or wash or endogenously to a plant to stimulate boththe immune responsiveness and growth characteristics of the plant thatcollectively result in improved yield performance. They can also provideprotection and growth benefits to the different parts of the plant (forexample, leaves, roots, tubers, corms, rhizomes, bulbs, pseudobulbs,flowers, pods, fruits, and growing meristems).

The combined foliar application or sequential applications of PSKα withHpaG-like bioactive priming polypeptides can be useful for enhancinggrowth of plants under standard (non-stress or optimal growth)environments or of plants exposed to abiotic stress (for example, heat,and water deficit stress).

Foliar application treatments using the X.spp HpaG-like and the At.PSKαbioactive priming polypeptides have different modes of action whenapplied on plants in optimal (non-stress) and in stress environments.The two classes of bioactive priming polypeptides are useful eitherprovided sequentially or in combination in a foliar application and canimprove plant growth in an environment that is with or without abioticstress(es).

X.spp.HpGa-like provides a plant growth benefit to corn in a non-stressenvironment where temperature, water, nutrients and other environmentalparameters were conducive to optimal plant growth. On the other hand,At.PSKα applied as a foliar spray provides a benefit to plant growthunder environmental conditions of heat and drought or water deficitstress. Thus, when used in combination in formulation together as foliarapplications they can span both non-stress and stress environments andprovide additive benefits to the growth of corn plants grown in avariety of environmental conditions.

Increases in plant productivity and growth for At.PSKα is also seen insoybean plants grown in environments with and without abiotic stress.Soybean plants that receive a foliar application with a formulationcontaining the bioactive priming polypeptide At.PSKα and are grown underconditions of heat and drought stress have increased yield over controlsoybean plants that received water and surfactant with no bioactivepriming polypeptide.

When X. spp. HpaG-like and At.PSKα are applied as a foliar spraytogether, they are useful to provide synergistic effects for plantproduction under normal and stressed environments. At.PSKα exhibitsincreased overall growth in corn when applied as a spray application,whereas X. spp. HpaG-like polypeptide results in the opposite trend.Thus, applying the two bioactive priming polypeptides together can actto balance plant growth in “heat stressed” environments such that thechanges in plant growth compared to control plants are greater than thesum of the effects of the bioactive priming polypeptides appliedindividually.

The synergistic interaction of these two classes of bioactive primingpolypeptides enhance plant growth under heat stressed environments(e.g., greater growth rates with increased plant biomass).

Any of the bioactive priming polypeptides as described herein can beapplied one or more times to a plant either in combination orindividually to enhance growth and productivity of a plant. Multipleapplications can be applied to promote yield benefits over the growingseason with applications tailored to the conditions in the environment,for example if a period of hot and dry weather is expected during thegrowth season, an additional spray of bioactive priming polypeptidesthat promote growth under abiotic stress can alleviate negative impactsto the plant.

Foliar Application of Phytosulfokine alpha (PSKα) to Increase Yield

A method is provided for applying At.PSKα as a foliar application toactively growing soybean plants to provide a yield advantage inenvironments with heat and drought stress. For example, a means ofapplying a composition containing bioactive priming At.PSKα polypeptideis provided as a foliar spray to soybean at V1-V4 stage usingapplication methods as described herein. Soybean plants treated withfoliar applications of At.PSKα can be grown in field environments underconditions that produced a non-stress and stress (heat and waterdeficit) environments. Treatment with At.PSKα can result in growth andyield benefits in plants grown in a variety of environmental conditionsincluding abiotic stressors.

Any of the RHPP bioactive priming polypeptides provided in Tables 12-14can be applied as a foliar, in furrow, seed treatment or root drenchapplication to a plant surface.

Foliar application of RHPP results in the alteration of plantarchitecture.

A method is provided where the RHPP polypeptide is applied as a foliarapplication to plants and results in a distinct leaf architecture (corn)and an enhanced root system (soybean). The increase in leaf angle androot biomass using a foliar treatment with RHPP has impactful advantagesfor use in agriculture in two major agriculture crops (corn andsoybean).

Application of RHPP to Alter Plant Architecture

Applying the bioactive priming polypeptide, RHPP, as a foliarapplication to V5-V8 corn results in a distinct leaf architecturephenotype with an upright leaf orientation and more erect leaves. Thisis particularly relevant with higher planting densities used to maximizeyield in a field environment. Foliar applications of the RHPPpolypeptide in maize (corn) is useful for changing the leaf angle thuscontributing to a smaller leaf angle which results in an upright leaforientation. This phenotype can be beneficial for increasing the leafarea index, reducing maize shade syndrome, and improving photosyntheticefficiency. In addition, providing RHPP as a foliar formulation tomaximize canopy development and total light penetrance is key toincreasing vegetative growth of the plants prior to the initiation ofthe grain filling stage.

Maize plants exhibit leaf curl or changes in their leaf architecture toa more upright leaf orientation to conserve water and enhance planttolerance to drought and heat. The upright changes to the leaf phenotypefor corn after application with the RHPP bioactive primingpolypeptide(s) compositions are useful and provide an alternativenon-breeding approach for shaping leaf architecture and enhancingtolerance to drought and heat.

An upright leaflet orientation phenotype in corn plants functions in thereduction of leaf temperatures, whole plant transpiration and in theimprovement of water use efficiency, as well as provide architecturalchanges to the plant canopy which can allow for higher density plantingsthat result in substantial increases in yield.

Application of the bioactive priming polypeptide, RHPP, to soybeans canalso provide benefits. For example, foliar application of RHPP toflowering soy can increase pod set. Pod set is a stage in soybeandevelopment occuring from the middle of R4 to the middle of R5 thatcontributes directly to yield. Initial pod set is marked by theemergence of a ¾ inch pod at one of the four uppermost nodes on the mainstem. It then progesses to the full pod stage where pod growth is rapidand seed development begins. An increase in pod set is quantified by anincrease in yield (i.e the pod number per node on a plant or the overallnumber of pods per plant).

RHPP to Increase Root Biomass and Yield

Soybean plants treated with foliar applied RHPP (SEQ ID NO: 600)bioactive priming polypeptide(s) can exhibit increased pod filling and amore-complete pod filing compared to non-treated plants which can be theresult of increases in nitrogen fixation.

Root architecture, particularly a root system with a rapid exploitationof deep soil can optimize nitrogen capture and water uptake which isespecially important in drying and nitrogen depleted soils. An RHPPpolypeptide(s) as described herein when applied as a foliar treatment tosoybean plants results in a root phenotype that is useful for water andmineral (nitrogen) acquisition, especially in nitrogen-deficient soils.Increasing nutrient uptake efficiency by enhancing root architecture isa key factor for improving plant productivity when used with soybeancultivation practices in a wide range of soil types.

Enhanced root biomass that results from a foliar application of RHPPprovided at the early vegetative stages for soybean VE-V5 or V2-V3 stageof development results in a root system with rapid exploitation of deepsoil (deep roots), and greater overall increases in root biomass. Forexample, a root hair promoting bioactive priming polypeptide such asRHPP (SEQ ID NO: 600) can be applied as a foliar treatment to soybeanplants at the V2 to V3 stage of development to result in an overallincrease in root biomass. Other notable enhancements in addition to rootbiomass are the production of longer lateral roots, increases in rootbranching, root hairs and increases in the root absorptive surface area.

RHPP can be applied as a foliar treatment at key developmental stages(VE-V8 or V2-V8) or in environments where a rapid increase in rootproduction is desired, such as dry or nutrient poor soil types. Soiltypes in particular may affect root development and expansion. Forexample, if plants have a hard time emerging in a clay soil, it mayaffect root formation and root proliferation. Increasing root mass maynot only beneficially effect plant emergence but also contribute toplant establishment. In addition, nodule formation and number areimportant because the bacteria that inhabit the nodules pull nitrogenfrom the air allowing soybeans to convert it into the nitrogen that theyneed to grow and produce seeds.

The RHPP polypeptides (Tables 13-15) can be used to increase noduleformation and nodule production of soybean roots when applied using anyof these treatment application methods which can be applied directly tothe soil, as a soil drench, as an in furrow treatment, or as a foliarapplication to the above ground plant parts.

Increase in nodules can result in increased nitrogen fixation bynitrogen fixing bacteria that inhabit the root nodules, such asRhizobium leguminosarum or japonicum. Nodule formation can be seenshortly after VE and can increase nitrogen fixation. Effectivenodulation of soybean roots results in higher yields and higher qualityseed production, protein and oil per seed or acre basis. Soybean plantshave fully formed first trifoliate leaves at the V1-V2 stage ofdevelopment which is estimated to be the peak time for nitrogenfixation.

The combination application of Gm.RHPP bioactive priming polypeptidewith various fertilizer treatment(s) can provide a yield boost and isrecommended especially for crop management applications in nitrogendepleted soils.

Bacterial Disease

Methods of using the bioactive priming polypeptides such as theflagellin-associated polypeptides or the thionin-like polypeptides asdescribed herein are useful for the prevention, treatment and control ofbacterial diseases in corn and particularly useful for the treatment ofbacterial leaf streak disease in corn caused by Xanthomonas vasicola pv.vasculorum, also recognized as Xanthomnas campestris pv. vasculorum.

Surveys indicate that bacterial leaf streak disease has spread and maybe widely distributed throughout the U.S. Corn Belt (Western Indiana,Illinois, Iowa, Missouri, Eastern Nebraska and Eastern Kansas). Diseasespread is most prevalent where corn is planted on corn in crop rotationpractices. The bacterial leaf streak disease can cause infection on dentcorn (field) seed corn, popcorn and sweet corn. The symptoms on corninclude narrow to brown yellow streaks and brown yellow strips betweenthe leaf veins. Lesions usually develop on lower or older plant leavesand initially spread to the higher or younger leaves on the plant.Yellow discoloration also may be present around lesions.

The bacterial leaf streak disease of corn presumably survives inpreviously infected host debris. Bacterial exudates found on surfaces ofinfected leaf tissues can serve as secondary inocula. The bacterium isspread by wind, splashing rain, and possibly by irrigation water. Thepathogen penetrates corn leaves through natural openings such asstomata, which can result in a banded pattern of lesions occurringacross leaves. Colonization of leaf tissues apparently is restricted bymain veins.

Because the disease is caused by a bacterial pathogen, the current useof bactericides is problematic to control it. For example, mostbactericides act as contact products and are not systemic and thus theywill not be absorbed or taken into the plant via other mechanisms.Bactericide treatments may require repeated applications as thebactericide may be washed off with rain or wind, thus rendering themuneconomical or impractical for use in some corn crops.

Current disease management practices to date recommend crop rotationpractices (such as corn, soybean and then back to corn) and theimplementation of sanitation practices, such as cleaning equipmentbetween field usage to slow disease progression.

Foliar applications of the Flg (Tables 4-5) and thionin polypeptides(Table 19) or combinations of the two classes provide an alternativeapproach for treating the disease. Foliar applications with thesebioactive priming polypeptides provided as a spray to the leaf surfaceof either asymptomatic or symptomatic plants provides a means toprevent, treat, and control the bacterial leaf streak disease in corn.

Alternatively, the flagellin- and thionin bioactive priming polypeptidesor combinations thereof can be useful for the prevention, treatment andcontrol of other bacterial diseases that infect corn (Table 21).

TABLE 21 Bacteria causing diseases in corn Corn Disease Bacteria CausingDiseases Bacterial leaf blight and stalk rot Pseudomonas avenae subsp.avenae Bacterial leaf spot Xanthomonas campestris pv. holcicolaBacterial leaf streak Xanthomonas vasicola Bacterial stalk rotEnterobacter dissolvens; Erwinia dissolvens Bacterial stalk and top rotErwinia carotovora subsp. carotovora Erwinia chrysanthemi pv. zeaeBacterial stripe Pseudomonas andropogonis Chocolate spot Pseudomonassyringae pv. coronafaciens Goss's bacterial wilt and blight Clavibactermichiganensis subsp. (leaf freckles and wilt) nebraskensis;Corynebacterium michiganense pv. nebraskense Holcus spot Pseudomonassyringae pv. syringae van Hall Purple leaf sheath Hemiparasitic bacteriaSeed rot-seedling blight Bacillus subtilis Stewart's disease (bacterialwilt) Erwinia stewartii Corn stunt (achapparramiento, Spiroplasmakunkelii maize stunt, Mesa Central or Rio Grande maize stunt)

Cercospora Leaf Blight Disease of Soybean

Cercospora is a fungal pathogen that causes the disease Cercospora leafblight of soybean. Cercospora leaf blight also referred to as the purpleseed stain disease infects both the leaves and seeds of soybeans.Cercospora infection of soybean seeds diminishes seed appearance andquality. The causal organism of Cercospora leaf blight is Cercosporakikuchii, which overwinters in soybean residue and in the seed coats.Spread of the disease occurs when the spores from the fungus are spreadto soybean plants from infected residue, weeds or other infected soybeanplants. Disease spread and symptom development are accelerated duringperiods of warm and wet weather. Symptom development usually beginsafter flowering and appears as circular lesions on soybean leaves asreddish brown to purple spots that can merge to form lesions. Symptomsare apparent in the upper canopy, usually in the uppermost three or fourtrifoliate leaves. Infected soybean plants exhibit worsening symptoms asthe crop matures, and premature defoliation of affected leaves may occurduring pod-fill. Cercospora symptom development may also appear aslesions on stems, leaf petioles and pods. Seeds are infected through theattachment to the pod. Cercospora infected seeds show a purplediscoloration, which can appear as specks or blotches covering theentire seed coat.

Foliar applications of flagellin or flagellin-associated polypeptides(Tables 4-5) provide an alternative approach for treating the disease.Foliar applications with these bioactive priming polypeptides providedas a spray to the leaf surface of either asymptomatic or symptomaticplants provides a means to prevent, treat, and control Cercospora LeafBlight in soybeans. Foliar applications of Flg22 derived from Bacillusthuringiensis, particularly at high use rates (e.g. 4.0 Fl. oz/Ac), canprovide a means of managing early symptom development and providehealthier more vigorous soybean plants grown in field locations thathave been impacted by Cercospora.

Specific combinations of bioactive priming polypeptides that can beuseful for treating or reducing the symptoms of Cercospora include: aflagellin or flagellin-associated polypeptide having an amino acidsequence comprising SEQ ID NO 226, 751 or 752; an RHPP polypeptidehaving a sequence comprising SEQ ID NO: 600; or a combination of aflagellin associated polypeptide having an amino acid sequencecomprising any one of SEQ ID NOs 226, 751 and 572 and an RHPPpolypeptide haivng the amino acid sequence comprising SEQ ID NO: 600.

For example, a useful combination of bioactive priming polypeptides fortreating, or reducing the symptoms of cercospora on a plant or plantpart is a flagellin polypeptide having an amino acid sequence comprisingSEQ ID NO: 226 alone or in combination with an RHPP polypeptide havingan amino acid sequence comprising SEQ ID NO: 600. Additional treatmentscan further comprise a fungicide in combination with these bioactivepriming polypeptides.

Asian Soybean Rust Disease

Asian soybean rust is a fungal disease caused by Phakopsora pachyrhizi.Its etiology and symptoms are similar to Cercospora and the bioactivepriming polypeptide combinations useful for treating it are similar aswell. Specifically, combinations of bioactive priming polypeptides thatcan be useful for treating or reducing the symptoms of Asian soybeanrust include: a flagellin or flagellin-associated polypeptide having anamino acid sequence comprising SEQ ID NO 226, 751 or 752; an RHPPpolypeptide having a sequence comprising SEQ ID NO: 600; or acombination of a flagellin associated polypeptide having an amino acidsequence comprising any one of SEQ ID NOs 226, 751 and 572 and an RHPPpolypeptide having the amino acid sequence comprising SEQ ID NO: 600.

For example, a useful combination of bioactive priming polypeptides fortreating, or reducing the symptoms of Asian soybean rust on a plant orplant part is a flagellin polypeptide having an amino acid sequencecomprising SEQ ID NO: 226 alone or in combination with an RHPPpolypeptide having an amino acid sequence comprising SEQ ID NO: 600.Additional treatments can further comprise a fungicide in combinationwith these bioactive priming polypeptides.

Holcus Spot

Holcus spot is a bacterial disease caused by Pseudomanas syringae pv.actinidae. Methods are described herein for using flagellin or flagellinassociated polypeptides to restrict growth of P. syringae and thusprevent or treat the disease of Holcus spot in a plant or a plant part.Flagellin or flagellin associated polypeptides useful for the treatmentof P. syringae include any polypeptides having amino acid sequencescomprising any one of SEQ ID NOs: 226, 540, 751, and 572 or anycombination thereof.

Sclerotinia Stem Rot (White Mold) Disease

Sclerotinia sclerotiorum is a plant pathogenic fungus that causes adisease caused white mold. It is also known as cottony rot, water softrot, stem rot, drop, crown rot, and blossom blight. Diagnostic symptomsof the white rot include black resting structures known as sclerotia andwhite fuzzy growths of mycelium on the infected plant. The sclerotia, inturn, produce a fruiting body that produces spores in a sac. Sclerotiniacan affect herbaceous, succulent plants, particularly fruits andvegetables, or juvenile tissue on woody ornamentals. It can also affectlegumes or tuberous plants like potatoes. White mold can affect a hostat any stage of growth, including seedlings, mature plants, andharvested products. It is usually found on tissues with high watercontent and close proximity to soil. Left untreated, pale to dark brownlesions on the stem at the soil line are covered by a white, fluffymycelial growth. This affects the xylem which leads to chlorosis,wilting, leaf drop, and death. White mold can also occur on fruit in thefield or in storage and is characterized by white fungal myceliumcovering the fruit and its subsequent decay. Flagellin or flagellinassociated polypeptides useful for the treatment of S. sclerotioruminclude any polypeptides having amino acid sequences comprising any oneof SEQ ID NOs: 226, 540, 571, 751, and 752.

Pseudomonas Leaf Spot

Pseudomonas syringae pv. actinidiae (PSA) is a devastating plantpathogen causing bacterial canker of both green- (Actinidiae deliciosa)and yellow-flesh (Actinidiae chinesis) kiwi plants throughout zones ofkiwi production, causing severe harvest loss in New Zealand, China, andItaly. In New Zealand alone, cumulative revenue losses to the mostdevastating biovar PSA-V are predicted to approach $740 million NewZealand leaves Dollars (NZD) by 2025 (Agribusiness and EconomicsResearch Institute of Lincoln University “The Costs of Psa-V to the NewZealand Kiwifruit Industry and the Wider Community”; May 2012). PSA-Vcolonizes the outer and inner surfaces of the kiwi plant and can spreadthrough the xylem and phloem tissues. Disease symptoms of PSA-V on kiwiinclude bacterial leaf spot, bacterial canker of the trunk, redexudates, blossom rot, discoloration of twigs, and ultimately dieback ofkiwi vines. The standard method of control for PSA-V currently employsfrequent foliar applications of metallic copper to kiwi vines which ispredicted to lead to the selection of copper-resistant form of thepathogen and loss of disease control. Novel methods of control areurgently needed.

Flagellin or flagellin associated peptides useful for the treatment ofPseudomanas syringase, particularly in kiwis, include any polypeptideshaving amino acid sequences comprising SEQ ID NO: 226, 540, 752, and/or571.

Asian Citrus Greening (Huanglonging) Disease

The methods described herein incorporate a different approach tocombating disease and additionally providing benefits of increasing theoverall productivity of a plant. This approach is specifically directedto providing either exogenous or endogenous applications of thebioactive priming polypeptides that include thionins to combat diseasein plants.

The thionin and thionin-like polypeptides (Table 19) and compositionsthereof are useful for the prevention, treatment and control of Asiancitrus greening also referred to as Huanglonging (HLB) disease, adevastating disease for citrus. HLB disease is widely distributed andhas been found in most commercial and residential sites in all countiesthat have commercial citrus orchards.

Methods are described herein for using the thionin polypeptides (SEQ IDNOs: 650-749) to prevent the spread of and in the treatment of HLBdisease.

Asian citrus greening disease is transmitted by the Asian citruspsyllid, Diaphorina citri or the two-spotted citrus psyllid, Triozaerytreae Del Guercio, which are both characterized as sap-sucking, hemipteran bug(s) in the family Psyllidae and have been implicated in thespread of citrus greening, a disease caused by a highly fastidiousphloem-inhabiting bacteria, Candidatus liberibacter asiaticus (Halbert,S. E. and Manjunath, K.L, “Asian citrus psyllids Sternorrhyncha:Psyllidae and greening disease of citrus: A literature review andassessment of risk in Florida,” Florida Entomologist 87: 330-353, 2004).Asian citrus greening or Huanglongbing disease is considered fatal for acitrus tree once the tree becomes infected.

The early symptoms of the disease on leaves are vein yellowing and anasymmetrical chlorosis referred to as blotchy mottle, which is the mostdiagnostic symptom of the disease. Infected trees are stunted andsparsely foliated with a blotchy mottling appearing on the foliage.Early symptoms of yellowing may appear on a single shoot or branch andwith disease progression, the yellowing can spread over the entire tree.Afflicted trees may show twig dieback, and fruit drop. Fruit are oftenfew in number, small, deformed or lopsided and fail to color properly,remaining green at the end and display a yellow stain just beneath thepeduncle (stem) on a cut fruit.

The Asian citrus greening disease may also be graft transmitted whencitrus rootstocks are selected for and grafted to scion varieties.

Management of citrus greening disease has proven difficult and thereforecurrent methods for control of HLB have taken a multi-tiered integrateddisease and pest management approach using 1) the implementation ofdisease-free nursery stock and rootstock used in grafting, 2) the use ofpesticides and systemic insecticides to control the psyllid vector, 3)the use of biological control agents such as antibiotics., 4) the use ofbeneficial insects, such as parasitic wasps that attack the psyllid, and5) breeding for new citrus germ plasm with increased resistance to thecitrus greening causing bacteria (Candidatus liberibacter spp.). The useof cultural and regulatory measures to prevent the spread of the diseaseis also part of the integrated management approach. Many aspectsinvolved in the management of citrus greening are costly both monetarilyand in respect to losses in citrus production.

Interveinal application of a thionin polypeptide or mixture of thioninpolypeptides can be delivered directly into the phloem (e.g., phloemcells including phloem sap, phloem companion cells and phloem sieve tubeelements) where Candidatus liberibacter can reside.

The thionins can be produced using an expression system where they canbe fused to a phloem targeting sequence(s) (Table 18) and then uniquelydelivered to the same vicinity where the bacteria can reside in thecitrus plant.

The phloem targeted thionin bioactive priming polypeptides are usefulfor treating citrus plants to prevent, reduce or eliminate the spread ofthe Asian citrus greening disease or Huanglonging (HLB) by directlytargeting the bacterium, Candidatus liberibacter asiaticus

These phloem targeted thionins can be delivered by injection into thephloem of a shrub or tree. Additionally, they can be delivered byspraying, washing, or adding as a soak or a drench to the soil or areasurrounding a plant.

Any of the phloem targeting sequences (Table 18; SEQ ID NOs: 641-649)can be used in combinations with the thionin and thionin-likepolypeptides (Table 19; SEQ ID NOs: 650-749).

The bacteria that cause HLB, Candidatus liberibacter asiaticus isdifficult to isolate and culture. In order to test individual thioninsand thionins with the phloem targeting sequences to determine if theyare useful for the treatment of HLB disease, Agrobacterium tumefacienscan be used as a model organism to test the effectiveness on reducingthe cell titer or growth of Agrobacterium prior to using the thionin orthionin combinations in an orchard setting.

The “peptide priming” methods provided herein with the thionins and/orthionin-like polypeptides (Table 19) can also be used in combinationflagellin and flagellin-associated polypeptides (Tables 1-5).Combinations of the thionin- and flagellin-associated bioactive primingpolypeptides can be used to prophylactically pre-treat a citrus plant byapplying the bioactive priming polypeptide or a composition containingthe polypeptide prior to the onset or appearance of anyinfection-related symptoms on the citrus shrubs or trees. Thispretreatment increases resistance to the disease pathogen that causescitrus greening (Candidatus liberibacter spp.).

The thionins provided in combination with the flagellin associatedbioactive priming polypeptides provide a more comprehensive approach todisease prevention and management. The thionin and flagellin associatedbioactive priming polypeptides use two distinct modes of action toprevent disease and the spread of disease.

The thionin-flagellin bioactive priming polypeptide combinations canalso be used with any other integrated management approach for diseasecontrol prescribed for HLB including, but are not limited to, (1) theuse of disease-free nursery stock and/or rootstocks for grafting, (2)the use of pesticides and/or systemic insecticides to control thedisease-causing psyllid, (3) the use of biological control agents suchas injections of antibiotics or parasitic insects that controls thepsyllid, (4) breeding new varieties of citrus germplasm with increasedresistance to the bacteria responsible for Asian citrus greeningdisease, (5) controlling parasitic plants (for example, dodder) that mayspread the disease, or (6) any combination thereof.

A synthetic version of a phloem targeting polypeptide (SEQ ID NO: 641)is particularly useful in targeting anti-microbial polypeptides to thephloem sieve tube and companion cells and can be useful for treatingvarious bacterial diseases of plants, such as bacterial leaf streak,Asian citrus greening or Huanglonging and citrus canker.

In addition, flagellin or flagellin associated polypeptides are usefulfor treating Asian citrus greening, especially when used in combinationwith a bacteriocide. For instance, flagellin or flagellin associatedpolypeptides having amino acid sequences comprising any one of SEQ IDNOs: 226, 571, and 752 can be used. Preferably, the bacteriocidecomprises oxytetracycline.

Citrus Canker

“Peptide priming” methods were developed for use with the bioactivepriming thionin and flagellin-associated polypeptides as described inTable 19 (thionins) and Tables 1-5 (flagellin and flagellin-associatedpolypeptides) to prophylactically treat citrus plants prior to anyvisible symptoms of the citrus canker disease or as a treatment once theonset of disease symptoms become apparent.

Citrus canker occurs primarily in tropical and sub-tropical climates andhas been reported to occur in over thirty countries including spread ofinfection reported in Asia, Africa, the Pacific and Indian OceansIslands, South America, Australia, Argentina, Uruguay, Paraguay, Braziland the United States. Citrus canker is a disease caused by thebacterium, Xanthomonas axonopodis pv. citri or pv. aurantifolii (alsoreferred as Xanthomonas citri subsp. citri) that infect foliage, fruitand young stems. Symptoms of citrus canker infection on leaves, andfruit of the citrus shrubs/trees can result in leaf-spotting, leaflesions, defoliation, die back, deformation of fruit, fruitrind-blemishing, pre-mature fruit drop, and canker formation on leavesand fruits. Diagnostic symptoms of citrus canker include acharacteristic yellow halo that surrounds the leaf lesions and awater-soaked margin that develops around the necrotic tissue on theleaves of the citrus plant. The citrus canker pathogen can spreadthrough the transport of infected fruit, plants, and equipment.Dispersal can also be facilitated by the wind and rain. Overheadirrigation systems may also facilitate movement of the citrus cankercausing pathogen. Infected stems can harbor the citrus canker causingbacteria (Xanthomonas axonopodis pv. citri) in the stem lesions fortransmission to other citrus plants. Insects, such as the Asianleafminer (Phyllocnistis citrella) also disemminate the disease.

In general, citrus plants susceptible to the citrus canker diseaseinclude orange, sweet orange, grapefruit, pummelo, mandarin tangerine,lemon, lime, swingle acid lime, palestine sweet lime, tangerine,tangelo, sour orange, rough lemon, citron, calamondin, trifoliate orangeand kumquat. World-wide, millions of dollars are spent annually onprevention, sanitation, exclusion, quarantine and eradication programsto control citrus canker (Gottwald T. R. “Citrus Canker,” The AmericanPhytopathological Society, The Plant Health Instructor 2000/updated in2005). Treatment for the disease has included application of antibioticsor disinfectants, the use of copper-based bactericidal sprays, andpesticide applications for Asian leafminer control.

The bioactive priming polypeptide combination comprising the thionin andthe flagellin-associated polypeptides can be applied to a citrus plantor citrus plant part (e.g., rootstock, scion, leaves, roots, stems,fruit, and foliage) using application methods that can comprise:spraying, inoculating, injecting, soaking, infiltrating, washing,dipping and/or provided to the surrounding soil as an in furrowtreatment.

The methods are provided using the bioactive priming polypeptidescomprising the thionin and/or flagellin-associated polypeptides topre-treat citrus plants or citrus plant parts (e.g., root stock, scion,leaves, roots, stems, fruit, and foliage) prior to any visibleoccurrence of symptoms. They are also useful for providing an increasein resistance to the citrus canker pathogen resulting in a reduction indisease symptoms.

Additionally, the methods of using the bioactive priming polypeptidessuch as the flagellin and flagellin-associated polypeptides are usefulto treat citrus plants or citrus plant parts (e.g., root stock, scion,leaves, roots, stems, fruit, and foliage) once the early onset of citruscanker disease symptoms or when the symptoms of the disease becomeapparent.

Application of the Flg polypeptides for treating citrus plants toprevent, reduce or eliminate the spread of the citrus canker disease canbe delivered by injecting into the phloem of a shrub or tree, spraying,washing, adding as a soak or a drench to the soil or soil areasurrounding a plant or provided in furrow.

Thionin bioactive priming polypeptides as described herein (Table 17)can be applied individually or in combination with any of theflagellin-associated Flg polypeptides (Tables 1-5) as a foliar treatmentor spray or as an injection and are useful for the prevention ofinfestation of citrus plants from insects such as the Asian leafminer(Phyllocnistis citrella) that have been identified in the disseminationof the bacteria (Xanthomonas axonopodis pv. citri) that cause the citruscanker disease.

Citrus Plants

Any of the methods described herein to provide improved plant health,disease tolerance or disease treatment applications to treat or preventAsian citrus greening (HLB) or citrus canker are suitable for use withany citrus plants and shrubs/trees.

The thionin or flagellin-associated polypeptides or compositionscomprising the thionin or flagellin-associated polypeptides as describedherein can be applied to any citrus shrub and/or tree and to anyagronomically-important citrus hybrid or citrus non-hybrid plant, andare useful for prophylactically treating the citrus to prevent the onsetof an infection or providing treatment after an infection has occurred.

Citrus plant species for use of the methods described herein include,but are not limited to: Sweet orange (Citrus sinensis, Citrus maxima xCitrus reticulata), Bergamot Orange (Citrus bergamia, Citrus limetta xCitrus aurantium), Bitter Orange, Sour Orange or Seville Orange (Citrusaurantium, Citrus maxima x Citrus reticulata), Blood Orange (Citrussinensis), Orangelo or Chironja (Citrus paradisi x Citrus sinensis),Mandarin Orange (Citrus reticulate), Trifoliate Orange (Citrustrifoliata), Tachibana Orange (Citrus tachibana), Clementine (Citrusclementina), Cherry Orange (Citrus kinokuni), Lemon (Citrus limon,Citrus maxima x Citrus medica), Indian Wild Orange (Citrus indica),Imperial Lemon (Citrus limon, Citrus medica x Citrus paradisi), Lime(Citrus latifoli, Citrus aurantifolia), Meyer Lemon (Citrus meyeri);hybrids of Citrus xmeyeri with Citrus maxima, Citrus medica, Citrusparadisi and/or Citrus sinensis), Rough Lemon (Citrus jambhiri),Volkamer Lemon (Citrus volkameriana), Ponderosa Lemon (Citrus limon xCitrus medica) Kaffir Lime (Citrus hystrix or Mauritius papeda), SweetLemon, Sweet Lime, or Mosambi (Citrus limetta), Persian Lime or TahitiLime (Citrus latifolia), Palestine Sweet Lime (Citrus limettioides),Winged Lime (Citrus longispina), Australian Finger Lime (Citrusaustralasica), Australian Round Lime (Citrus australis), AustralianDesert or Outback Lime (Citrus glauca), Mount White Lime (Citrusgarrawayae), Kakadu Lime or Humpty Doo Lime (Citrus gracilis), RusselRiver Lime (Citrus inodora), New Guinea Wild Lime (Citrus warburgiana),Brown River Finger Lime (Citrus wintersii), Mandarin Lime (Citruslimonia; (hybrids with Citrus reticulata x Citrus maxima x Citrusmedica), Carabao Lime (Citrus pennivesiculata), Blood Lime (Citrusaustralasica x Citrus limonia) Limeberry (Triphasia brassii, Triphasiagrandifolia, Triphasia trifolia), Grapefruit (Citrus paradisi; Citrusmaxima x Citrus xsinensis), Tangarine (Citrus tangerina), Tangelo(Citrus tangelo; Citrus reticulata x Citrus maxima or Citrus paradisi),Minneola Tangelo (Citrus reticulata x Citrus paradisi), Orangelo (Citrusparadisi x Citrus sinensis), Tangor (Citrus nobilis; Citrus reticulata xCitrus sinensis), Pummelo or Pomelo (Citrus maxima), Citron (Citrusmedica), Mountain Citron (Citrus halimii), Kumquat (Citrus japonica orFortunella species), Kumquat hybrids (Calamondin, Fortunella japonica;Citranqequat, Citrus ichangensis; Limequat, Citrofortunella floridana;Orangequat, hybrid between Satsuma mandarin x Citrus japonica orFortunella species; Procimequat, Fortunella hirdsiie; Sunquat, hybridbetween Citrus meyeri and Citrus japonica or Fortunella species;Yuzuquat, hybrid between Citrus ichangensis and Fortunella margarita),Papedas (Citrus halimii, Citrus indica, Citrus macroptera, Citrusmicrantha), Ichang Papeda (Citrus ichangensis), Celebes Papeda (Citruscelebica), Khasi Papeda (Citrus latipes), Melanesian Papeda (Citrusmacroptera), Ichang Lemon (Citrus ichangensis x Citrus maxima), Yuzu(Citrus ichangensis x Citrus reticulata), Cam sành (Citrus reticulata xCitrus maxima), Kabosu (Citrus sphaerocarpa), Sudachi (Citrus sudachi),Alemow (Citrus macrophylla), Biasong (Citrus micrantha), Sam uyao(Citrus micrantha), Kalpi (Citrus webberi), Mikan (Citrus unshiu),Hyuganatsu (Citrus tamurana), Manyshanyegan (Citrus mangshanensis), Lush(Citrus crenatifolia), Amanatsu or Natsumikan (Citrus natsudaidai),Kinnow (Citrus nobilis x Citrus deliciosa), Kiyomi (Citrus sinensis xCitrus unshiu), Oroblanco (Citrus maxima x Citrus paradisi), Ugli(Citrus reticulata x Citrus maxima and/or Citrus x paradisi), Calamondin(Citrus reticulata x Citrus japonica), Chinotto (Citrus myrtifolia,Citrus aurantium or Citrus pumila), Cleopatra Mandarin (Citrus reshni),Daidai (Citrus aurantium or Citrus daidai), Laraha (Citrus aurantium),Satsuma (Citrus unshiu), Naartjie (Citrus reticulata x Citrus nobilis),Rangpur (Citrus limonia; or hybrid with Citrus sinensis x Citrus maximax Citrus reticulata), Djeruk Limau (Citrus amblycarpa), Iyokan,anadomikan (Citrus iyo), Odichukuthi (Citrus odichukuthi), Ougonkan(Citrus flaviculpus), Pompia (Citrus monstruosa), Taiwan Tangerine(Citrus depressa), Shonan gold (Citrus flaviculpus or Citrus unshiu),Sunki (Citrus sunki), Mangshanyen (Citrus mangshanensis, Citrusnobilis), Clymenia (Clymenia platypoda, Clymenia polyandra), Jabara(Citrus jabara), Mandora (Mandora cyprus), Melogold (Citrus grandis xCitrus paradisii/Citrus maxima/Citrus grandis), Shangjuan (Citrusichangensis x Citrus maxima), Nanfengmiju (Citrus reticulata), andShīkwāsaī (Citrus depressa).

The thionin and/or flagellin-associated priming polypeptides can beapplied to any citrus plant, shrub/tree used for medicinal orcosmetic/health and beauty purposes, such as Bergamot Orange (Citrusbergamia), Sour or Bitter Orange (Citrus aurantium), Sweet Orange(Citrus macrophylla), Key Lime (Citrus aurantiifolia), Grapefruit(Citrus paradisi), Citron (Citrus medica), Mandarin Orange (Citrusreticulate), Lemon (Citrus limon, or hybrids with Citrus medica x Citrusmaxima, Citrus limonia, Citrus medica x Citrus maxima x Citrus medica),Sweet Lime (Citrus limetta), Kaffir Lime, (Citrus hystrix or Mauritiuspapeda), Lemon hybrid or Lumia (Citrus medica x Citrus limon), (Citrusmedica x Citrus maxima x Citrus medica), Omani Lime (Citrusaurantiifolia, Citrus medica x Citrus micrantha), Jambola (Citrusgrandis), Kakadu Lime or Humpty Doo Lime (Citrus gracilis), Pomelo(Citrus retkulata), Tangor (Citrus nobilis), and Sour Lime or Nimbuka(Citrus acida).

Exemplary important citrus hybrids for fruit production are: SweetOrange (Citrus sinensis), Bitter Orange (Citrus aurantium), Grapefruit(Citrus paradisi), Lemon (Citrus limon), Persian Lime (Citruslatifolia), Key Lime (Citrus aurantiifolia), Tangerine (Citrustangerine) and Rangpur (Citrus limonia).

Additionally, any of the bioactive priming polypeptides, compositions,and methods as described herein can be applied to any citrus plant,shrub/tree used as a rootstock and/or a scion germplasm. The methods areparticularly useful for rootstocks commonly used in grafting of citrusto enhance the merits of the scion varieties, which can includetolerance to drought, frost, disease or soil organisms (for example,nematodes). Such citrus plants that provide useful rootstocks include:Sour or Bitter Orange (Citrus aurantium), Sweet Orange (Citrusmacrophylla), Trifoliate Orange (Poncirus trifoliata), Rough Lemon(Citrus jambhiri), Volkamer Lemon (Citrus volkameriana), Alemow (Citrusmacrophylla), Cleopatra Mandarin (Citrus reshini), Citrumelo (hybridswith x citroncirus species), Grapefruit (Citrus paradisi), Rangpure Lime(Citrus limonia), Palestine Sweet Lime (Citrus limettioides) and TroyerCitrange (Citrus sinensis x Poncirus trifoliata or Citrus sinensis xCitrus trifoliata) and Citrange (Citrus sinensis x Poncirus trifoliataor C. sinensis x C. trifoliata).

Use of Retro-Inverso Flg Bioactive Priming Polypeptides to Treat andReduce Citrus Greening

Combinations of flagellin-associated polypeptides paired with theirretro-inverso counterparts can be used to treat and reduce the greeningeffect on citrus that results in Asian citrus greening or Huanglongbingdisease.

An early symptom of HLB in citrus is the yellowing of leaves on anindividual limb or in one sector of a tree's canopy. Leaves that turnyellow from HLB will show an asymmetrical pattern of blotchy yellowingor mottling of the leaf, with patches of green on one side of the leafand yellow on the other side. As the HLB disease progresses, the fruitsize becomes smaller, and the juice turns bitter. The fruit can remainpartially green and tends to drop prematurely.

Treatment combinations of Flg polypeptides with their retro-inverso (RI)forms can be used to minimize the effect on citrus fruit greening. Suchtreatment combinations can be applied on HLB-infected trees. Theretro-inverso forms will compete with the native forms of Flgpolypeptides for binding to the FLS-associated receptor(s) at the plantsurface and thus inhibit/delay the symptom formation of greeningassociated with HLB disease. The native Flg22 and RI Flg22 combinationsassist with a fine tuned immune response to reduce and even eliminatethe disease-causing bacteria, Candidatus liberibacter asiaticus, whilepreventing acute symptom development, such as leaf yellowing and citrusfruit greening.

EXAMPLES

The following non-limiting examples are provided to further illustratethe present invention.

Example 1: Application of Bt.4Q7Flg22 and Retro-Inverso Bt.4Q7Flg22, andEc.Flg22 and Ec.RI Flg22 to Corn

The effect of Bt.4Q7Flg22 (SEQ ID NO: 226) and retro-inverso Bt.4Q7Flg22(SEQ ID NO: 376), as well as Ec. Flg22 (SEQ ID NO 526) and Ec.RI Flg22(SEQ ID 527) bioactive priming polypeptides on corn (BECK'S 5828 YH,6175YE) yield was determined in 10 separate locations in the US Midwest(FIG. 2 and FIG. 3).

Field seed beds at each location were prepared using conventional orconservation tillage methods for corn plantings. Fertilizer was appliedas recommended by conventional farming practices and remained consistentbetween the US Midwest locations. Herbicides were applied for weedcontrol and supplemented with cultivation when necessary. Four-rowplots, 17.5 feet (5.3 meters) long were planted at all locations. Cornseed was planted 1.5 to 2 inches (3.8 to 5.1 cm) deep, to ensure normalroot development, at 28,000 to 36,000 plants per acre with row widths of30 inch (76.2 cm) rows with seed spacing of approximately 1.6 to 1.8seeds per foot. Each hybrid was grown in at least three separate plots(replicates) at each location to account for field variability.

Native Bt.4Q7Flg22 bioactive priming polypeptide (SEQ ID NO: 226) andits retro-inverso polypeptide (SEQ ID NO: 376) were chemicallysynthesized via solid phase peptide synthesis and formulated at 0.33 Fl.oz/Ac (24.1 mL/hectare, Ha) use rate. The final concentration in thespray tank was 25 nM after dilution in carrier rate of 10 gallonswater/Ac (37.85 L/Ha). Native Bt.4Q7Flg22 bioactive priming polypeptideswere applied during first and second year field trials to measureeffects across a multi-year growing season. Retro-inverso polypeptideswere applied during the first year field trials to compare with nativeBt.4Q7Flg22. Bioactive priming polypeptides were applied as foliar sprayapplications at 0.33 Fl. oz/Ac (24.1 mL/Ha) use rate during the V5-V8development stage. Each polypeptide was applied with a non-ionicsurfactant at 0.5%. The effect of bioactive priming polypeptides wasmeasured as the absolute changes in yield in bushels per acre (Bu/Ac).Additionally, the win rate was calculated: the percentage of testinglocations at which one treatment has a yield advantage over othertreatments (in this case, as compared to the non-treated controlplants).

FIG. 2, panel A shows that during the first year field trials, foliarspray application of Bt.4Q7Flg22 (SEQ ID NO: 226) resulted in an averageyield increase of 11.60 Bu/Ac (728.1 kg/Ha) and a win rate of 90% acrossthe 10 locations compared to the non-treated control corn plants. FIG.2, panel B shows that foliar spray applications of retro inversoBt.4Q7Flg22 bioactive priming polypeptide (SEQ ID NO: 376) resulted inan average yield increase of +11.90 Bu/Ac (746.9 kg/Ha) and a win rateof 70% across the 10 locations in the US Midwest compared to thenon-treated control corn plants. In both figures, locations (1-12) arereported on the x-axis and absolute change in yield Bu/Ac is reported onthe y-axis and above or below the bar graphs at each location. Ec. Flg22polypeptide delivered to corn yielded 8.2 bu/Ac (514.7 kg/Ha) advantagewith a 80% win rate across the 10 sites. The retro inverso version ofEc.RI Flg22 did not yield as well, giving 1.9 bu/c across the 10 siteswith a 50% win rate.

The second year field trials were conducted using large acre fieldtrials at 10-11 locations in the US MidWest (IL, IN, IA) and employedfoliar spray application of the Bt.4Q7Flg22 bioactive primingpolypeptide (SEQ ID NO: 226) provided to V8 corn plants (Dekalb 5064).Foliar spray application of Bt.4Q7Flg22 was applied at a use rate of0.33 fluid ounces per acre (Fl. oz/Ac). As shown in FIG. 3, foliarapplication using the Bt.4Q7Flg22 bioactive priming polypeptide resultedin an average yield increase of +4.8 Bu/Ac over the control across the11 locations with a win rate of 83%. Locations 1-6 are reported on thex-axis and absolute change in yield Bu/Ac is reported on the y-axis andabove or below the bar graphs at each location.

First year field trials using foliar treatments using Bt.4Q7Flg22bioactive priming polypeptide (SEQ ID NO: 226) applied to corn hybrid(BECK's 5828 YH) shown in FIG. 2 (panel A) resulted in over a +11 Bu/Ac(690.4 kg/Ha) increase in yield over the non-treated control plants.Second year field trials applied to V8 corn plants (Dekalb 5064) shownin FIG. 3 resulted in an almost+5 Bu/Ac (313.8 kg/Ha) increase comparedto the yield of the non-treated control plants. The combined average forthe two corn hybrids resulted in a 2-year average yield increase of +8.0Bu/Ac (50.2 kg/Ha) across locations with a win rate of 86% representedfor the multiple year growing season.

A third study was performed with Bt.4Q7 Flg22 bioactive primingpolypeptide tested as a V5-V8 application to corn at 3 rates and appliedwith a non-ionic surfactant. The final use rates were 0.33 Fl. oz/Ac, 4Fl. oz/Ac, 8 Fl. oz/Ac (24.1 mL/Ha, 292.3 mL/Ha, 584.6 mL/Ha), resultingin approximate final concentrations of 25 nM, 300 nM, 600 nMrespectively. Each study was performed at between 10 and 11 sites. Theend results of the study at V5-V8 at 0.33 Fl. oz/Ac (24.1 mL/Ha) was5.75 Bu/Ac or 360.9 kg/Ha advantage, at 4 Fl. oz/Ac a 3.77 bu/Ac or236.6 kg/Ha advantage, and at 8 Fl. oz/Ac a 5.05 Bu/Ac or 317 kg/Ha.

Example 2: Application of Bt.4Q7Flg22 to V8 Corn with Fungicide

Foliar treatments with Bt.4Q7Flg22, with and without a commerciallyavailable fungicide, STRATEGO YLD, were conducted to determine ifsynergistic effects resulted from the combinations of the Bt.4Q7Flg22bioactive priming polypeptide with the fungicide. Foliar sprayapplication of Bt.4Q7Flg22 (SEQ ID NO: 226) alone or in combination withSTRATEGO YLD was assessed on corn plants (hybrid Dekalb 5064) at the V8stage of development.

Replicated trials were conducted at 6-8 locations throughout the USMidwest (IA, IL, IN) using replicated trials. Corn plants were grown asdescribed in Example 1. Plots were maintained using the individualgrower's production practices and each plot was replicated 3-4 times.When used, STRATEGO YLD fungicide (a combination of prothioconazole andtrifloxystrobin) was applied using the recommended label rates (4.0Fl.oz/Ac or 292.3 mL/Ha)) at each location. Foliar treatmentapplications consisted of the following treatments: (a) non-treatedcontrol, (b) STRATEGO YLD fungicide alone, and Bt.4Q7Flg22 (SEQ ID NO:226) delivered in a free peptide form provided with (c) and without (d)the fungicide. Bt.4Q7Flg22 was applied at a use rate of 0.33 or 4.0fluid ounces per acre (Fl. oz/Ac) or (24.1 or 292.3 mL/Ha).

Corn yield in bushels per acre (Bu/Ac) was reported at all locations asan average yield for the replicated trials at each location. The changein yield in Bu/Ac for corn plants receiving foliar applications with theSTRATEGOYLD fungicide were normalized to the average yield for thecontrol corn plants for the 6 locations (Table 22).

Foliar treatments with Bt.4Q7Flg22 provided at 0.33 Fl. oz/Ac (24.1mL/Ha) provided yield benefits over the non-treated control corn plantswith a +4.84 Bu/Ac (303.8 kg/Ha) increase observed across the 6locations. Foliar treatment using only the fungicide application ofSTRATEGO YLD also provided a yield benefit in corn of +4.88 Bu/Ac (306.3kg/Ha) over the control plants. Application of the free peptide,Bt.4Q7Flg22, at 0.33 Fl. oz/Ac (24.1 mL/Ha) combined with STRATEGO YLDfungicide at 4.0 Fl. oz/Ac demonstrated a synergistic effect, resultingin an average of +10.72 Bu/Ac (672.9 kg/Ha) over the non-treated controlplants. Therefore, the Bt.4Q7Flg22 polypeptide and fungicide treatmentcombination resulted in a synergistic effect at the 0.33 Fl. oz/Ac (24.1mL/Ha) use rate for the polypeptides and 4.0 Fl. oz/Ac (292.3 mL/Ha) userate for the fungicide.

TABLE 22 Foliar treatment of corn with Bt.4Q7Flg22 bioactive primingpolypeptide applied with a fungicide to increase yield in corn AverageTotal Average Bu/Ac Application Yield Increase Use Rate Bu/Ac comparedto Treatment - Corn Fl. oz/Ac (6 locations) Control Control — 187.37 —Bt.4Q7Flg22 0.33 192.21 +4.84 STRATEGO YLD 4.0 193.80 +4.88Bt.4Q7Flg22 + 0.33 207.86 +10.72 STRATEGO YLD 4.0

A second study looking at the combination of Ec. Flg22 with STRATEGO wasalso performed at 8 sites as replicated trials in the same fashion asabove. Ec. Flg22 at 4 Fl. oz/Ac (292.3 mL/Ha) added 1.3 (Bu/Ac) (81.6kg/Ha) on top of STRATEGO YLD with a 63% win percentage over 8 sites.This demonstrates that both Flg22 polypeptides were able to add benefitover a commercial fungicide, STRATEGO YLD.

Example 3: Application of Bt.4Q7 Flg22, Retro-Inverso Bt.4Q7Flg22,Ec.Flg22, Retro Inverso Ec.Flg22 or RHPP to R2 Soybean—Increased Yield

Foliar application using, Bt.4Q7 Flg22 bioactive priming polypeptide(SEQ ID NO: 226; FIG. 4, panel A), the retro-inverso (RI) Bt.4Q7Flg22(SEQ ID NO: 376; FIG. 4, panel B) from Bacillus thuringiensis strain 4Q7and root hair promoting polypeptide (RHPP, SEQ ID NO: 600) derived fromGlycine max were applied individually to soybean plants (commercialhybrid Beck's 294 NR) at the R2 stage of development using a use rate of0.33 Fl. oz/Ac or 24.1 mL/Ha (Flg22 polypeptides) or 4.0 Fl. oz/Ac or292.3 mL/Ha (RHPP). Cultivation methods employed in Example 1 werefollowed in growing soybean seeds. Soybean seed (commercial hybridBeck's 294 NR) was planted 1.5 to 2 inches (3.8 to 5.1 cm) deep toassure normal root development. Soybean seed was planted atapproximately on average 150,000 plants per acre with row widths of 30inch (76.2 cm) rows with seed spacing of approximately 7 to 8 seeds perfoot (0.3 meter).

Yield results in bushels per acre (Bu/Ac) are reported for soybean grownin 11 separate US Midwest locations harvested in October (FIG. 4).Soybean yield (Bu/Ac) is also reported as averaged across all of thelocations as the change in yield (Bu/Ac) normalized to the controlsoybean plants. Soybean yield following foliar application withBt.4Q7Flg22 (SEQ ID NO: 226) and the RI Bt.4Q7Flg22 (SEQ ID NO: 376) wascompared to yield of non-treated soybean plants and plotted in FIG. 4.Locations 1-11 are reported on the x-axis. Absolute change in yield(Bu/Ac) as compared to the non-treated control soybean plants isreported on the y-axis and above or below the bar graphs at eachlocation. Average yield across all 11 locations are reported andhighlighted with the black bar. Soybean yield for the Bt4Q7Flg22 and RIBt.4Q7Flg22 foliar treated plants showed similar trends at the 11different locations. Spray application using RI Bt.4Q7Flg22 on soybeanresulted in an average yield increase of 0.90 Bu/Ac (60.5 kg/Ha) for the2 soybean hybrids across 11 locations compared to the soybeannon-treated control plants. Yield results for the natural (all L)Bt.4Q7Flg22 in soybean was neutral (−0.1 Bu/Ac or −6.7 kg/Ha)) whencompared across the locations. Yield data represented across 11individual US Midwestern locations resulted in a win rate of 64%, forboth the RI Bt.4Q7Flg22 and Bt.4Q7Flg22 spray application treatments ascompared to the control or non-treated soybean plants. The addition ofRHPP polypeptide at 4 Fl. oz/Ac (292.3 mL/Ha) in the same studyincreased yield by 1.2 Bu/Ac (80.7 kg/Ha) compared to control.

A second study was performed to test Ec. Flg22 and Ec.RI Flg22polypeptides as R2 foliar treatments on soybeans with a carrier rate of10 gallons/Ac (93.5 L/Ha) water and NIS surfactant. A concentration of100 nM was obtained in the tank for each treatment. The application ofthe Ec. Flg22 lead to a 0.9 Bu/Ac (60.5 kg/Ha) increase with a 82% winrate for the 11 sites, and the Ec.RI Flg22 lead to a 0.6 Bu/Ac (40.3kg/Ha) increase with a 80% win rate over 10 sites. Also included was theRHPP polypeptide as a seed treatment, with 1.2 bu/Ac (80.7 kg/Ha) over11 sites at 73% win rate.

A third study at the same 11 sites was performed adding a foliarfertilizer alone or with RHPP at 8 fl oz/Ac (584.6 mL/Ha). The additionof RHPP on top of the foliar fertilizer gave 1 Bu/AC (67.2 kg/Ha)advantage across the 11 sites.

Example 4: Foliar Spray Application of Flg22 Polypeptides to Soybeans

Foliar treatments with Bt.4Q7Flg22, Ec.Flg22, and RHPP at 4.0 and 8.0Fl. oz/Ac (292.3 and 584.6 mL/Ha) were tested at the R2 timing onsoybean varieties over 11 sites with 10 gallons/Ac (93.5 L/Ha) waterwith 0.5% NIS surfactant. At the higher dose of 8 Fl. oz/Ac (584.6mL/Ha), the Ec. Flg22 polypeptide gave a 0.74 Bu/Ac (49.8 kg/Ha)advantage and the Bt.4Q7 Flg22 gave a 0.88 Bu/Ac (59.2 kg/Ha) advantage.The lower rate of 4 Fl. oz/Ac (292.3 mL/Ha) for RHPP gave 0.31 Bu/Ac(20.9 kg/Ha) yield advantage.

Example 5: Application of Escherichia coli Flagellin Polypeptides toIncrease Yield—Corn

The effect of flagellin polypeptides derived from Escherichia coli oncorn yield was then tested. Corn plants (Beck's 5828 YH) received aninitial spray application at the V5-V8 stage of development withformulations containing the Ec. Flg22 bioactive priming polypeptide (SEQID NO: 526) and the retro-inverso RI Ec.Flg22 (SEQ ID NO: 527) fromEscherichia coli applied at a use rate of 0.33 Fl. oz/Ac (24.1 mL/Ha).Yield results in bushels per acre (Bu/Ac) were determined for corn grownin the 12 separate locations harvested in October.

FIG. 5 depicts yield across these 12 locations, normalized to thenon-treated control plants and shown as an increase or a decrease inBu/Ac compared to the control. Yield data represented for 12 individuallocations in Illinois resulted in a win rate of 50%. Corn plants thatreceived a foliar spray application of Ec.Flg22 bioactive primingpolypeptide (FIG. 5, panel A) resulted in an average yield increase of+8.2 Bu/Ac (514.7 kg/Ha) across the 12 locations over non-treatedplants. Corn plants that received the foliar spray applications of theretro inverso RI Ec.Flg22 bioactive priming polypeptide (FIG. 5, panelB) resulted in an average yield increase of +1.9 Bu/Ac 119.3 kg/Ha)across the 12 locations as compared to the non-treated control cornplants. Therefore, application of foliar sprays containing the Ec.Flg22(SEQ ID NO: 526) bioactive priming polypeptides provided a beneficialgrowth response and yield benefit to corn plants when applied at theV5-V8 stage of development.

Example 6: Foliar Application of Escherichia coli Flagellin Polypeptidesto V2-V3 Soybean to Increase Plant Height

Foliar application of the Ec.Flg22 (SEQ ID NO: 526) and retro inverso RIEc.Flg22 (SEQ ID NO: 527) was applied to soybean (Beck's 297NR). Plantswere grown in an environmentally controlled growth room. Seed wasplanted directly into 39.7 cm³ pots containing Timberline top soil at adepth of 2.54 cm, with 2 seeds per pot. After planting, 50 mL of roomtemperature water was added to each pot to allow for germination. Thepots were kept in an artificial lighted growth room receivingapproximately 300 μmol m⁻² s⁻¹ (light photons) for a 13/11 light/daycycle and a 21° C. day/15° C. night temperature range. Plants receivedthe same watering and fertilizer regimes.

Foliar treatments using both the native and retro inverso forms ofEc.Flg22 were applied to 3-week-old soybean plants at the V2 to V3 stageof development using a use rate of 0.33 Fl. oz/Ac (24.1 mL/Ha). Plantheight (cm) was measured just prior to the foliar application deliveredat 3 weeks and then again 2 weeks later when the plants were5-weeks-old. Two replicate trials were conducted using 18 plants pertrial.

As described in Table 23, foliar application of the Ec.Flg22 polypeptideto soybean at the V2-V3 stage of development increased plant height,compared to the control (water only treatment) plants (Table 23). Foliarapplication using the Ec.Flg22 (SEQ ID NO: 526) and the retro-inversoEc.Flg22 (SEQ ID NO: 527) bioactive priming polypeptides resulted in+13% and +16% increases in plant height when normalized to the controlnon-treated soybean plants (normalized to 100%).

TABLE 23 Foliar application of flagellin polypeptide increases plantheight for soybean Percentage Foliar Height (cm) Height (cm) height ofTreatment at 3 weeks at 5 weeks control Soybean Ec.Flg22 (1 μM) 40.17(5.83) 64.79 (8.40) 113.2% Ec.Flg22-Retro 36.57 (6.00) 66.46 (5.77)116.1% Inverso (1 μM)

Example 7: Application of Flg22 and Retro Inverso Flg22 in Corn—PlantHeight

Corn (Beck's hybrid 5828 YH) plants were grown in an environmentallycontrolled growth room as described in Example 6. Plants were measuredthree weeks after emergence and then treated with foliar applications ofnatural (L) and retro-inverso (D) forms of Flg22 polypeptides fromBacillus thuringiensis (Bt.4Q7Flg22, SEQ ID Nos 226 and 376) andEscherichia coli (Ec.Flg22, SEQ ID NOs: 526-527). Bioactive primingpolypeptides were applied as free polypeptides at a concentration of 1μM. Control plants were treated with water alone. After an additional 2weeks of growth, plant height was measured (at 5 weeks).

The change in plant height (Δ height cm) between the 2 week and 5 weekinterval time points was measured and normalized to the growth ofwater-treated control plants. Three replicate trials were conductedusing 9 plants per trial equaling a total of 27 measurements pertreatment (Table 24). There were no differences in the plant heightmeasured between the EcFlg22, the Bt.4Q7Flg22 or the water treatedcontrol plants at the 3-week measurement time point. The greatest changein plant height from 3 to 5 weeks was reported for corn plants thatreceived the Ec.Flg22 foliar application (Δ=17.60 cm). These plants alsoachieved a +8.3% increase in height compared to control plants at the 5week measurement mark. The two retro inverso polypeptides (RI Ec.Flg22and RI Bt.4Q7Flg2) and the natural Bt.4Q7Flg22 similarly increased plantheight when compared to the control treatment with increases reportedfrom approximately +2% to +4%.

TABLE 24 Foliar application of Ec.Flg22 and Bt.Flg22 polypeptides oncorn resulted in increases in plant height Δ Height Height HeightNormalized as Foliar (cm) at (cm) at a percentage Treatment 3 weeks 5weeks Δ Height of control Corn (STDEV) (STDEV) (cm) height Ec. Flg2247.00 (8.30) 64.60 (6.93) 17.60 +8.3% 1 μM Ec. Flg22 48.62 (6.62) 62.00(4.07) 13.38 +3.9% Retro inverso 1 μM Bt 4Q7Flg 22 50.10 (6.79) 61.89(7.03) 5.40 +3.7% 1 μM Bt 4Q7Flg22 49.05 (4.28) 61.03 (7.13) 11.98 +2.3%Retro inverso 1 μM

Example 8: Application of Retro Inverso Flg22 Bioactive PrimingPolypeptides to Promote Growth Under Stress—Corn

Abiotic stress causes significant crop loss and can result in majorreductions in crop production and yield potential. The flagellincompositions and flagellin-associated bioactive priming polypeptides canbe used as chemical priming agents to increase tolerance of a plant toone or more abiotic stresses. Foliar treatments using the Ec.Flg22 andBt.4Q7Flg22 and the retro inverso (RI) forms of both of these bioactivepriming polypeptides were conducted to determine if these foliar appliedpolypeptides could provide a protective advantage against heat anddrought stress.

Corn (Beck's hybrid 5828 YH) seed was planted and grown as described inExample 6 with the difference that a 16 hour day/8 hour nightlight-cycle was followed. Temperature was cycled from 21° C./day to 15°C./night with 75% humidity. The light cycle still provided a uniformapproximately 300 μmol m⁻² s⁻¹, adequate light for plant growth. Plantswere measured at 3 weeks after emergence and were then treated withfoliar applications of natural or the retro-inverso (RI) forms ofEc.Flg22 (SEQ ID NOs 526-527) or Bt.4Q7Flg22 (SEQ ID NOs: 226 and 376)at 1 μM concentrations. Control plants were treated only with water. Aweek after the spray treatments were applied, the plants were subdividedinto 2 groupings where one group remained in the same standard growthenvironment as described and the other group was transferred to anenvironment that provided heat and water deficit stress. In the heatstress environment, the temperature was elevated using heat maps from21° C. to 27° C. for 18 hours per day for a period of 5 days. Plantswere left un-watered for the heat stress duration to further simulate awater deficit stress. Change in plant height (cm) was measured 2 weekslater at 5 weeks and reported as a percentage of the height of thecontrol (water) plants. Measurements are reported as the combinedaverage of two trials with 9 replicate plants per trial (Table 25).

As shown in Table 25, both natural forms of the bioactive primingpolypeptides (Ec.Flg22 and Bt.4Q7Flg22) increased plant growth, asmeasured by control plant height, when applied under non-stressedconditions. The two treatments resulted in plants that reached heights103% and 108% of their respective controls. However, only corn plantstreated with the retro inverso Flg22 polypeptides (both retro inversoEc.Flg22 and Bt.4Q7Flg22) showed enhanced plant growth compared tocontrol plants when grown in both normal and heat/water stressedenvironments. Plants treated with Ec. Flg22 Retro inverso reached 103%of their control heights in both conditions. Plants treated withBt.4Q7Flg22 reached 102% and almost 108% of their counterpart control'sheights in non-stressed and stressed conditions, respectively.

Therefore, corn plants that were treated with the retro inverso Flg22polypeptides (RI-Ec.Flg22 and RI-Bt.4Q7Flg22) exhibited increased growthas indicated by increased percentage in plant height over the controlplants. This result suggests that the retro-inverso forms are morestable in form and able to survive without proteolytic breakdown inharsher environments or situations conducive to abiotic stress. Thus,they may offer a protective advantage to plants that are subjected toabiotic stress environments.

TABLE 25 Foliar application of Ec.Flg22 and Bt.Flg22 to corn grown innon-stress and stress environments Non-Stressed Δ Height Stressed ΔHeight Foliar Treatment (cm) Normalized as a Normalized as a in Corn(Non-heat percentage of control percentage of control stressed) heightheight Ec. Flg22 1 μM 108.3% 95.8% Ec. Flg22 Retro 103.9% 103.5% inverso1 μM Bt. 4Q7Flg 22 103.7% 100.1% 1 μM Bt.4Q7Flg22 Retro 102.3% 107.8%inverso 1 μM

Example 9: Heat and Water Deficit Stress after Application of FoliarFlg22 Polypeptide to V2-V3 Corn

In a separate experiment, corn plants, grown as described in Example 8,were treated with Bt.4Q7Flg22 along with a surfactant before exposure toheat and water deficit stress. Three replicate trials of 18 corn plantreplicates per trial were grown in an environmentally controlled growthroom until the V2-V3 stage of development. Each plant was treated withfoliar sprays containing 0.1% surfactant with or without Bt.4Q7Flg22 (1μM final concentration). A week after the spray treatments were applied,the plants were transferred to an environment that provided a heatstress and water deficit stress. Heat stress was applied using heat matsto raise the temperature in the environment from 21° C. to 27° C. Duringthe period of heat stress, the plants were left unwatered. The cornplants remained in the simulated abiotic stress environment for one weekand then plant height (cm) was re-measured (Table 26).

As shown in Table 26, in two out of the three trials, application ofBt.4Q7Flg22 polypeptide applied as a foliar spray (Trials 1 and 3)resulted in significant increased growth (height measured in cm) in cornplants as compared to the control plants treated with the surfactantalone. Foliar treatment with the Bt.4Q7Flg22 bioactive primingpolypeptide resulted in an almost 13% increase in plant height in Trial1 and more than a 33% increase in Trial 3 compared to the control(surfactant alone treated) plants.

TABLE 26 Change in plant height in corn with application of Bt.4Q7Flg22Δ Height Height (cm) Height (cm) Normalized as before stress afterstress a percentage Treatments 2 weeks 4 weeks Δ Height of control Corn(STDEV) (STDEV) (cm) height Trial 1 Surfactant 17.62 (2.32) 27.96 (3.02)+10.34 100.0% (0.1%) Bt.4Q7Flg22 17.72 (2.08) 29.39 (3.04) +11.68 112.9%(1 μM) Trial 2 Surfactant 15.93 (1.22) 25.26 (1.99) +9.32 100.0% (0.1%)Bt.4Q7Flg22 16.03 (1.97) 25.31 (6.29) +9.28 99.5% (1 μM) Trial 3Surfactant 13.16 (2.28) 21.43 (2.89) +8.28 100.0% (0.1%) Bt.4Q7Flg2214.99 (1.97) 26.02 (3.21) +11.03 133.2% (1 μM)

Example 10: Seed Treatment Using the Flg22 Polypeptides—Corn and Soy

Corn seed from two separate hybrids (hybrid BECK's 5828 AM and 4606 P2)was treated with Bt.4Q7Flg22 (SEQ ID NO: 226) bioactive primingpolypeptides with final slurry concentrations of 0.25 μM or 1.0 μM(Table 27) applied to the surface of each seed. The seed applicationswere provided using a 40 μM polypeptide stock diluted to the appropriateconcentration in a slurry containing a fungicide, insecticide,beneficial bacteria, colorant and seed finisher (EverGol Energy (0.031mg ai/seed), PONCHO/VOTiVO (0.6 mg ai/seed), Peridium 1006 (5 fl oz/cwtor 147.9 mL/cwt) and Pro-Ized Red Colorant (normal) (0.5 fl oz/cwt).Seed treatment was applied using a Wintersteiger HEGE II (WintersteigerAG, Austria, Germany).

Seed was planted in 12 locations in the U.S. Midwest (IA, IL, IN).Sixteen randomized replicate blocks were harvested per each of the Flg22polypeptide treatments consisting of Bt.4Q7Flg22 applied at 0.25 μM and1.0 μM slurry concentration.

Table 27 shows that seed treatment with the Bt.4Q7Flg22 bioactivepriming polypeptide applied at what would be the equivalent of a 40 μMpolypeptide solution at a rate of 0.035 or 0.14 Fl. oz (2.6 or 10.2mL/Ha) of polypeptide solution per unit of corn seed resulted inenhanced yield with averages of +2.1Bu/Ac (131.8 kg/Ha) increases forthe low rate and +5.3 Bu/Ac increases for the high rate application ascompared to non-treated control seed (no seed treatment).

TABLE 27 Seed treatment on corn using Flg22 polypeptides EquivalentAverage Change in Peptide Application Average Average Total Yield YieldBu/Ac concentration Rate Total Yield Total Yield Bu/Ac compared toTreatment in seed Fl. oz/unit Bu/Ac Bu/Ac Hybrid 1 the Control Corncoating slurry corn seed Hybrid 1 Hybrid 2 and 2 Seed Control — — 206.65184.27 197.32 — Bt.4Q7Flg22 0.25 μM 0.035 fl oz 218.36 182.88 200.62+2.1 of 40 μM peptide solution/ unit Bt.4Q7Flg22  1.0 μM 0.14 fl oz213.44 187.48 202.62 +5.3 of 40 μM peptide solution/ unit

A second study was set up to test the ability of Ec.Flg22, Ec.RI Flg22,Bt.4Q7Flg22, and Bt.4Q7RI Flg22 to promote yield in corn. Replicatedtrials with 12 locations were set up as above. The Bt.4Q7 Flg22 gave 2.8bushels or 71.1 kg at 50% win rate, the Bt.4Q7 RI Flg22 polypeptide gave0.5 Bu/Ac. The Ec. Flg22 polypeptide gave 2.8 Bu/Ac (175.8 kg/Ha)advantage at 70% win rate, and the Ec.RI Flg22 gave no benefit.

A third study was set up to look at soybean seed treatment benefits ofBt.4Q7 Flg22, RI Bt.4Q7Flg22, Ec.RI Flg22, and RHPP as a seed treatmenton soybean. Over a 12 location study, the RHPP polypeptide gave 0.4Bu/AC (26.9 kg/Ha) at 64% win rate, the Bt.4Q7 Flg22 polypeptide gave1.3 Bu/Ac (87.4 kg/Ha) at 64%, the Bt.4Q7 RI Bt.4Q7Flg22 polypeptidegave 0.3 Bu/Ac (20.2 kg/Ha) at 55%, and the Ec.RI Flg22 gave 1.8 Bu/Ac(121.1 kg/Ha) at 73% win rate.

Example 11: Application of Flagellin Bioactive Priming Polypeptides toTomatoes—Increased Yield

Foliar application treatments of Bt.4Q7 Flg22 (SEQ ID NO: 226) and Ec.Flg22 (SEQ ID NO: 526) were applied as an exogenous spray at thepre-bloom stage and used to increase yield in tomatoes.

Small scale plots were designed to simulate commercial growingconditions for tomatoes. Two hybrids of tomatoes, JetSetter (Trial 1)and Better Big Boy (Trial 2) were started as transplants in thegreenhouse 42 to 56 days prior to planting in the raised field beds.Tomatoes were transplanted once soil temperatures three inches (7.6 cm)beneath the soil surface reach 60° F. (15.5° C.). Tomatoes were grown onraised beds covered with black plastic mulch. Plants were grown usingdrip irrigation and fertilizer applied following grower guidelinesthroughout the growing season to ensure optimum plant growth and yields.Small raised bed plots were designed to simulate the planting densitiesused by commercial growers that generally plant 2,600 to 5,800 plantsper acre in single rows with 18 to 30 inches (46 to 76 cm) betweenplants in the row on 5- to 6.5-ft (1.5 to 2 m) centers.

Foliar treatments of Bt.4Q7 Flg22 and Ec. Flg22 at low and high userates of 1 Fl. oz/Ac (73.1 mL/Ha) and 20 Fl. oz/Ac (1461.5 mL/Ha),respectively, were applied on the two hybrids at early bloom (firstflower) stage. Replicated trials were conducted at the University ofMissouri (Columbia, Mo.) in July. Control plants were treated with equalvolumes (use rates) of water. Effects of the foliar treatments onincreasing yield in tomatoes were determined and reported as normalizedto the water control treatment. The average percentage change in yieldover the average control yield is reported in the Table 28.

Foliar application of both the Bt.4Q7Flg22 and Ec.Flg22 bioactivepriming polypeptides increased tomato fruit yield for each hybrid atboth the low and high use rate. When results for the two hybrids wereaveraged, low and high application use rates for Bt.4Q7 Flg22 increasedtomato yield+25% and +17%, respectively, over the control plants.Similarly, low and high application use rates for the Ec. Flg22treatments resulted in an average increase in tomato yield of +43% and+46% over the control plants for the two hybrids.

TABLE 28 Foliar treatment to increase yield in different hybrids oftomato Trial 1: Trial 2: Percent Percent Change in Change in AverageTrials Yield over Yield over 1 & 2 Avg. Control; Avg. Control; PercentChange Hybrid: Hybrid: Better Yield over Avg. Foliar Treatment JetsetterBig Boy Control Bt.4Q7 Flg22 +49%  +1% +25% (1 Fl. oz/Ac) Bt.4Q7 Flg22+22% +12% +17% (20 Fl. oz/Ac) Ec. Flg22 +61% +25% +43% (1 Fl. oz/Ac) Ec.Flg22 +72% +21% +46% (20 Fl. oz/Ac)

Example 12: Foliar Treatment of Tomato Plants with a Formulation ofBt.4Q7 Flg22

In another experiment, tomato plants (hybrid: Better Boy), cultivated asdescribed in the previous example, were treated with a formulation ofBt.4Q7 Flg22 at the first bloom stage. The formulation used consisted ofthe retro inverso D RI Bt.4Q7 Flg22 applied with 0.01% (v/v) non-ionicsurfactant. The formulation wasapplied to tomato foliage usingapplication use rates of 1 Fl. oz/Ac (73.1 mL/Ha) in two replicatewinter tomato trials conducted in Florida. At harvest, the yield wasmeasured as the number of fruits per plant, the weight (grams) per fruitand the total yield (lbs/Ac). Table 29 reports the yield as a percentcomparison or change to the non-treated control (water only) plants.

Foliar treatment using the Bt.4Q7 Flg22 formulation applied at 1 Fl.oz/Ac (73.1 mL/Ha) increased yield of Better Boy tomatoes an average of21% compared to the non-treated (water alone) control plants. Thisincrease for Better Boy tomatoes corresponded to both an increase innumber of fruits per plant and an increase in the fruit weight (Table29).

TABLE 29 Foliar treatment with a Flg22 bioactive priming polypeptide toincrease yield in tomato Percent Change in Number of Percent ChangePercent Change Fruits per Plant in Weight/Fruit in Yield (lbs/Ac)Compared to Compared to Compared to Treatment Control Control ControlBt.4Q7 Flg22 +12% +9% +21% 1 Fl. oz/Ac

Example 13: Application of Flagellin Bioactive Priming Polypeptides toPeppers—Increased Yield

Foliar treatments of Bt.4Q7 Flg22 (SEQ ID NO: 226) and Ec.Flg22 (SEQ IDNO: 526) were applied as an exogenous spray at the first-bloom stage andused to increase yield in two pepper varieties.

Foliar treatments of Bt.4Q7 Flg22 and Ec.Flg22 bioactive primingpolypeptides were applied using small scale plots designed to simulatecommercial growing conditions for peppers (Capsicum). Two varieties ofpepper: Red Knight (RK) and Hungarian Hot Wax (HHW) were grown from6-week old transplants in raised beds covered with black plastic mulchthat had good water-holding characteristics and a pH of 5.8-6.6. Plantswere grown using drip irrigation and fertilizer applied following growerguidelines throughout the growing season to ensure optimum plant growthand yields. Small raised bed plots were designed to simulate theplanting densities used by commercial growers that generally plantapproximately 10,000-14,000 plants per acre in double rows 14-18 inches(35.6 to 46 cm) apart on plastic mulched beds with 16-24 inches (40.6 to61 cm) between plants in the row and with the beds spaced 5.0-6.5 feet(40.6 to 70 cm) apart from their centers. A single row of peppers alsocan be planted on each bed (5,000-6,500 plants per acre or 12,355-16,062plants per hectare).

Foliar applications with compositions containing Bt.4Q7 Flg22 andEc.Flg22 were applied at the first flower stage at an application userate of 1 Fl. oz/Ac (low rate) or 73.1 mL/Ha and 20 Fl. oz/Ac (highrate) or 1461.5 mL/Ha on both pepper plants and compared to the control(water applied at same use rate). Effects of the foliar applications onpepper yield were determined for two separate harvests using a once overharvest approach and normalized to the yield of the control plants. Theaverage percentage change in yield for each treatment over the yield forthe control plants is reported as pounds/acre (lbs/Ac) in Table 29.

Foliar treatment of peppers using either the Bt.4Q7 Flg22 or Ec.Flg22bioactive priming polypeptides resulted in overall average increases inpepper yield (lbs/Ac) with both the low and high application use ratesand for both the RK and HHW pepper varieties. The combined yieldaverages for the RK and HHW varieties were +53% higher (low rate: 1 Fl.oz/Ac or 73.1 mL/Ha) and +25% higher (high rate: 20 Fl. oz/Ac) forBt.4Q7 Flg22 foliar treated peppers compared to the control pepperplants. Alternatively, the combined yield average increases for the RKand HHW varieties were +30% higher (low rate: 1 Fl. oz/Ac) and +47%higher (high rate: 20 Fl. oz/Ac or 1461.5 mL/Ha) for Ec. Flg22 foliartreated peppers compared to the control pepper plants.

Differences existed in how the two pepper varieties responded to thefoliar treatments and in the resultant yield advantages provided to bothpepper varieties (Table 30). Substantial yield increases were seen inthe HHW variety as compared to the RK variety of peppers and the controlor non-treated plants with yield increases of +77% (low: 1 Fl. oz/Ac or73.1 mL/Ha) and +42% (high: 20 Fl. oz/Ac or 1461.5 mL/Ha) over thecontrol or non-treated pepper plants for the Bt.4Q7Flg22. Additionally,low use rates of the Bt.4Q7Flg22 (1 Fl. oz/Ac) and high use rates of theEc. Flg22 (20 Fl oz/Ac) polypeptides were the most effective atincreasing yield in the HHW variety (both yielded a +72% increase overthe control plants).

TABLE 30 Foliar treatment of Flg22 to increase yield in differentvarieties of pepper Avg. Percent Avg. Percent Change Yield CombinedChange Yield Total Weight Avg. Percent Total Weight (lbs/Ac) ChangeYield (lbs/Ac) (Hungarian Total Number Red Knight Hot Wax) (lbs/Ac)Foliar Treatment 2 Replicate Trials 2 Replicate Trials RK and HHWBt.4Q7Flg22: +29% +77% +53% 1 Fl. oz/Ac Ec.Flg22: +30% +29% +30% 1 Fl.oz/Ac Bt.4Q7Flg22:  +8% +42% +25% 20 Fl. oz/Ac Ec.Flg22: +22% +72% +47%20 Fl. oz/Ac

Example 14: Application of Flagellin Bioactive Priming Polypeptides toSquash—Increased Yield

Foliar treatment of Bt.4Q7Flg22 were applied exogenously on Ambassadorsquash at the first bloom stage using two separate formulations(formulation 1=F1 and formulation 2=F2). Formulation 1 (F1) consists ofthe native L Bt.4Q7 Flg22 bioactive priming polypeptide applied with0.01% (v/v) non-ionic surfactant. Formulation 2 (F2) consists of the DRI Bt.4Q7 Flg22 applied with 0.01% (wv) non-ionic surfactant. Bothformulations F1 and F2 were applied to squash foliage using applicationuse rate of 1 Fl. oz/Ac (73.1 mL/Ha). Yield comparisons were madebetween the plants treated with the foliar Bt.4Q7Flg22 F1 and F2 sprayapplications compared to the control (water) or non-treated squashplants. Squash plants were cultivated in sandy loam soil as follows. 2.5cm holes were cut in 2.5 ft. (0.76 m) wide plastic covered mounds, tworows per mound, holes spaced 1.5 ft (0.46 m) apart within each row. Rowswere staggered within the mound. Mounds were spaced 4 ft (1.2 m) apart.Three squash seeds were planted per hole and thinned to a single plantper hole 14 days after planting. Drip irrigation tubing was laid in thecenter of each mound, and plants were watered as necessary.

Squash plants were grown from seed in raised beds until bloom, andfoliar treated in the same Florida (FL) location using two replicatedtrials or two separate harvests. Yield for the foliar Bt.4Q7Flg22applied F1 and F2 treated plants is reported as the number of squash perplant, the weight (grams) per squash and the total squash yield (lbs/Ac)and represented as a percentage change as compared to non-treatedcontrol plants (Table 31).

Foliar treatments of Bt.4Q7Flg22 using the two formulations F1 and F2resulted in an increased yield advantage when foliar applied on squash(Ambassador) at the pre-bloom stage compared to the non-treated controlplants. The number of squash per plant, weight per squash and overallaverage percent change in yield (lbs/Ac) all were increased in theBt.4Q7Flg22 F1 and F2 treated plants compared to the control ornon-treated plants. The squash plants treated with both the Bt.4Q7Flg22F1 and F2 formulations had similar trend increases in the number ofsquash per plant, weight per squash and overall average percent changein yield (lbs/Ac), however squash plants that received the F1 foliarapplication showed increases in the number of squash per plant and inthe total yield of squash over the plants that received the F2formulation.

TABLE 31 Foliar treatment with a composition of Flg22 polypeptides toincrease yield in squash Percent Change Percent Change in Number ofPercent Change in Yield Squash per in Weight/Squash (lbs/Ac) PlantCompared Compared to Compared to Treatment to Control Control ControlBt.4Q7Flg22 +7% +2% +9% 1 Fl. oz/Ac Formulation 1 Bt.4Q7Flg22 +4% +2%+6% 1 Fl. oz/Ac Formulation 2

Example 15. Screening Flg Polypeptides for Reactive Oxygen Species (ROS)Production in Corn and Soybean

Codon usage was performed to generate mutations in the Bt.4Q7Flg22 tobetter match the host organism and the binding of the Flg22 polypeptideto the FLS receptor at the plant cell surface. A probabilistic approachwas used to generate three variants of the native Bt.4Q7Flg22 that weredesigned to have preferred amino acid signatures for corn and soybeanand to perform equal to or better than the native Bt.4Q7Flg22(SEQ ID NO:226) in ROS activity assays. These variants possessed mutations to theinternal segment (SEQ ID NO: 571), or the C-terminus (SEQ ID NO: 572) orthe N terminus (SEQ ID NO: 573) and were designated asBt.4Q7Flg22-Syn01, Bt.4Q7Flg22-Syn02 and Bt.4Q7Flg22-Syn03,respectively. Bt.4Q7Flg22-Syn01 and Bt.4Q7Flg22-Syn03 were then measuredin relation to their native forms at a variety of concentrations.

Fresh plant tissues from corn (hybrid 5828 YX) and soybean (hybrid 297R4) leaves were cut into uniform samples and floated on 150 μL ofsterile water in a 96-well white, low luminescence plate. The plate wasplaced under growth lights that had a 16-hour light/8-hour dark cyclesat a consistent temperature of 22° C.

For corn samples, aerial tissue from V1 to V4 stage corn plants was cutaway from the plant above the soil line using a clean razor blade. Thecotyledon and sheath were removed. 1-mm slices were cut through thestalk from the base of the plant until approximately 1.3 cm below thefirst leaf node. Each corn section was placed in an individual well ofthe 96-well plate.

For soybean samples, fully expanded trifoliate leaves were removed fromV1-V3 stage plants. Leaf discs (12.6 mm²) were cut from the leaf bladesusing a 4-mm diameter clean, sharpened cork borer. Discs were cut inhalf using a clean razor blade, and each disc half was placed in anindividual well of the 96-well plate.

Native Flg22 polypeptide (SEQ ID NO: 226) or Flg22 polypeptidescontaining the described mutations (SEQ ID NOs 571 or 573) stocks wereprepared in either sterile, deionized water or 100 mM sodium phosphate(pH 7.8-8.0) buffer with 0.1% Tween-20. After 18-24 hours, the water wasremoved from each well of the 96-well plate. Plant tissue samples weretreated with a 100 μL elicitation solution containing 1:100 dilution ofFlg22 polypeptide stock (concentration range from 250 picomolar (μM) to10 micromolar (μM)), 34 μg/mL luminol, and 20 μg/mL horseradishperoxidase. Recognition of the Flg22 polypeptide by the plant tissueresulted in activation of immune signaling and the production ofapoplastic reactive oxygen species (ROS). In the presence of ROS (H₂O₂),horseradish peroxidase catalyzed the oxidation of luminol and productionof visible light. Relative light units (RLUs) were recorded with aGLOMAX 96 microplate luminometer (Promega Corporation) using a 0.5 sintegration; 2.6 min intervals over a time course of 40 minutes.

For data analysis, total RLUs produced were calculated for each sampleover the entire 40 min time course. Significant outliers beyond theinterquartile range were excluded from analysis. Total RLUs in eachcondition (n=6-16) were normalized to the average RLU for Bt.4Q7Flg22 at25 nM and reported as a percentage (%) of the Bt.4Q7Flg22 control (Table32).

The synthetic mutagenized Bt.4Q7Flg22-Syn01 version had increased ROSactivities at a range of concentrations (0.25-100 nM) whileBt.4Q7Flg22-Syn03 was more varied and showed increased ROS activities at0.25 nM, 1 nM, 10 nM, 25 nM, and 100 nM concentrations as compared tothe native version of Flg22 or Bt.4Q7Flg22. The synthetic version ofBt.4Q7Flg22-Syn01 treatment using 5 nM resulted in the largest change inROS activity over the native version or Bt.4Q7Flg22. ROS activities forBt.4Q7Flg22-Syn03 showed a more varied response over the range ofconcentrations added.

TABLE 32 Flg generated synthetic mutants (Syn-01 and Syn-03) have moreactivity in the ROS assay than the native Bt.4Q7Flg22 over a wide rangeof concentrations. Concentration Bt.4Q7Flg22 Bt.4Q7Flg22-Syn01Bt.4Q7Flg22-Syn03 (nM) (SEQ ID NO: 226) (SEQ ID NO: 571) (SEQ ID NO:573) 0.25 8.12 23.56 14.86 0.5 14.86 55.00 14.86 1 23.85 57.04 41.47 557.00 113.00 57.00 10 76.85 116.34 85.70 25 100.00 118.05 111.40 100113.74 120.83 162.29 1000 127.76 121.20 97.67

Example 16: ROS Screening Assays to Identify Functionally Active FlgPolypeptides for Corn and Soybean

Based on the results from preliminary studies in Example 15, thefollowing concentrations were chosen to screen ROS activities of a widerange of Flg22 polypeptides in corn and soybean: 5 nM in corn (hybrid5828 YX) and 100 nM in soybean (hybrid 297 R4). ROS activity assays werethen used to identify the best Flg22 bioactive priming polypeptidecandidates for individual treatment use of corn and soybean and toidentify those candidates that were active for both corn and soybean.

Corn and soybean leaf tissues were harvested from plants and ROS assayswere performed as previously described in Example 15 for the mutantpolypeptides listed in Table 33. Total RLUs produced were calculated foreach sample over the entire 40 min time course. Significant outliersbeyond the interquartile range were excluded from analysis. Comparisonsof ROS activity on corn (hybrid 5828 YX) and soybean (hybrid 297 R4)were made and reported as the percentage (%) of relative light units(RLU) compared to the average RLU values at the 25 nM Bt.4Q7Flg22treatment concentration (Table 33).

Table 33 summarizes the relative activity for a variety of mutant Flg22polypeptides compared to native Bt.4Q7Flg22 alongside the standarddeviation in for each condition (STDEV).

TABLE 33 ROS activity comparisons for variousFlg22 polypeptides in corn and soybean Corn (5828 YX) Soybean (297 R45 nM 100 nM polypeptide polypeptide Avg. Avg. Amino Acid ActivityActivity SEQ ID NO: Sequence (%) STDEV (%) STDEV Bt.4Q7Flg22DRLSSGKRINSA 100 — 100 — Bacillus SDDAAGLAIA thuringiensisSEQ ID NO: 226 Bt.Flg22-Syn01 DRLSSGKRINSA 142.9 39.3 112 3.0Mutant S13K KDDAAGLAIA Bacillus thuringiensis SEQ ID NO: 571Bt.Flg22-Syn02 DRLSSGKRINSA 78.3 26 68.7 14.0 Mutant A20Q SDDAAGLQIABacillus thuringiensis SEQ ID NO: 572 Bt.Flg22-Syn03 QRLSSGKRINSA 122.129.5 113.5 42.6 Mutant D1Q SDDAAGLAIA Bacillus thuringiensisSEQ ID NO: 573 Bm.Flg22-B1 NRLSSGKQINSA 106.0 25.2 74.6 4.9 BacillusSDDAAGLAIA manliponensis SEQ ID NO: 290 Ba.Flg22-B2 NRLSSGKRINSA 134.756.8 83.0 26.6 Bacillus ADDAAGLAIA anthracis SEQ ID NO: 295 Bc.Flg22-B3DRLSSGKRINNA 80.3 18.4 90.0 35.5 Bacillus cereus SDDAAGLAIASEQ ID NO: 294 A spp.Flg22-B4 ERLSSGYRINRA 78.1 20.1 133.1 23.9Aneurini-bacillus SDDAAGLAIS spp. XH2 SEQ ID NO: 300 Ba.Flg22-B5EKLSSGQRINSA 27.1 2.3 42.2 7.4 Bacillus SDDAAGLAIS aryabhattaiSEQ ID NO: 289 P spp.Flg22-B6 GKLSSGLRINGA 135.3 31.6 112.5 22.8Paenibacillus spp. SDDAAGLAIS strain HW567 SEQ ID NO: 293 L spp.Flg22-L1LRLSSGYRINSA 26.6 3.6 64.1 14.1 Lysinibacillus spp. ADDAAGLAISSEQ ID NO: 291 L spp.Flg22-L2 EKLSSGLRINRA 104.5 1.2 128.6 29.5Lysinibacillus spp. GDDAAGLAIS SEQ ID NO: 580 L spp.Flg22-L3EKLSSGYKINRA 36.4 7.9 96.9 20.6 Lysinibacillus spp. SDDAAGLAISSEQ ID NO: 581 L spp.Flg22-L4 LRISSGYRINSA 60.1 5.9 117.9 25.7Lysinibacillus spp. ADDPAGLAIS SG9 SEQ ID NO: 582 Lf.Flg22-L5LRISTGYRINSA 59.3 5.8 111.6 27.5 Lysinibacillus ADDPAGLAIS fusiformisSEQ ID NO: 583 Lm.Flg22-L6 EKLSSGFRINRA 58.7 19.4 112.3 42.3Lysinibacillus GDDAAGLAIS macroides SEQ ID NO: 584 Lm.Flg22-L6EKLSSGYKINRA 33.7 1.4 77.0 19.2 Lysinibacillus GDDAAGLAIS xylanilyticusSEQ ID NO: 585 Pa.Flg22 QRLSTGSRINSA 116.0 32.5 88.6 22.2 PseudomonasKDDAAGLQIA aeruginosa SEQ ID NO: 530 Ec.Flg22 ERLSSGLRINSA 95.0 46.7116.8 13.3 Escherichia coli KDDAAGQAIA SEQ ID NO: 586 Xcc.Flg22QRLSSGLRINSA 143.3 5.2 96.4 17.6 Xanthomonas KDDAAGLAIS campestris pvcampestris strain 305 or (Xanthomonas citri pv. citri) SEQ ID NO: 532Ea.Flg22 QRLSSGLRINSA 125.2 9.2 91.9 10.1 Erwinia amylovora KDDAAGQAISSEQ ID NO: 534 Bp.Flg22 TRLSSGKRINSA 111.2 14.0 67.2 3.0 BurkholderiaADDAAGLAIS phytofirmans strain PsJN SEQ ID NO: 536 Bu.Flg22 NRLSSGKRINTA92.9 12.7 91.1 12.9 Burkholderia ADDAAGLAIS ubonensis SEQ ID NO: 538Ps.Flg22 TRLSSGLKINSA 154.4 20.7 113.1 19.6 Pseudomonas KDDAAGLQIAsyringae pv. actinidiae ICMP 19096 SEQ ID NO: 540

Based on the results from Table 33, a number of predictions could bemade based on the effect of different mutations on Flg22 polypeptides onROS activity in corn and soybean. Table 34 describes ROS activityobserved or predicted for a variety of targeted mutations. Briefly,replacements at the first amino acid (D1N, D1Q or D1T) have or likelywill result in strong recognition and/or activation of the Flg22receptor in corn. Mutations in the inner segment, K7Y, K7F and A16P,will likely have similar positive results in soybean. Of the testedpolypeptides, Bt.4Q7Flg22-Syn01 (S13K) and Bt.4Q7Flg22-Syn03 (D1Q) hadthe strongest ROS-inducing activity in corn and soybean.

TABLE 34 Result summary of mutant versions of native Bt.4Q7Flg22Amino Acid Description of ROS SEQ ID NO Sequence activity resultsSEQ ID NO: 226 DRLSSGKRINS Bt.4Q7Flg22 ASDDAAGLAIABacillus thuringiensis (native version and  used as the standardcomparison) SEQ ID NO: 571 DRLSSGKRINS S13K mutation: Strong AKDDAAGLAIAROS activation in both corn and soybean SEQ ID NO: 572 DRLSSGKRINSA20Q mutation: Negative ASDDAAGLQIA ROS activation inboth corn and soybean SEQ ID NO: 573 QRLSSGKRINS D1Q mutation: StrongASDDAAGLAIA ROS activation in both corn and soybean SEQ ID NO: 574NRLSSGKRINS D1N mutation: Strong ASDDAAGLAIA ROS activation inboth corn and soybean* (predicted) SEQ ID NO: 575 TRLSSGKRINSD1T mutation: Strong ASDDAAGLAIA ROS activation inboth corn and soybean* (predicted) SEQ ID NO: 576 DRLSSGYRINSK7Y mutation: Strong ASDDAAGLAIA ROS activation in only soybean*(predicted) SEQ ID NO: 577 DRLSSGFRINS K7F mutation: Strong ASDDAAGLAIAROS activation in only soybean* (predicted) SEQ ID NO: 578 DRLSSGKRINSA16P mutation: Strong ASDDPAGLAIA ROS activation in only soybean*(predicted) SEQ ID NO: 579 DRLSSGKRINS K7Q mutation: Strong ASDDAAGLAIAreduction in ROS activation in both corn and soybean* (predicted)

Example 17: ROS Activity Assays to Identify Combinations of FlgPolypeptides for Corn and Soybean

Corn and soybean leaf tissues were harvested from plants and ROS assayswere performed as previously described in Example 15. The relative ROSactivity of different Flg22 variants, alone or in combination, wereassessed to identify the preferred combinations of Flg22 polypeptidesthat when applied together provided the highest ROS activity responsefor both corn and soybean. Results are summarized in Table 35.

TABLE 35 Flg22 combinations with increasedROS activities in corn and soybean Corn (5828 YX) Soybean (297 R4) 5 nM100 nM polypeptide polypeptide Flagellin Amino Acid Avg. Avg.Composition Sequence Activity (%) STDEV Activity (%) STDEV Bt.4Q7Flg22DRLSSGKRINS 100 — 100 — Bacillus ASDDAAGLAIA thuringiensisSEQ ID NO: 226 Bt.Flg22-Syn01 DRLSSGKRINS 122.48 31.69 83.54 36.21Bacillus AKDDAAGLAIA thuringiensis SEQ ID NO: 571 Ba.Flg22-B2NRLSSGKRINS 142.53 7.45 97.59 68.59 Bacillus antrhacis AADDAAGLAIASEQ ID NO: 295 A spp.Flg22-B4 ERLSSGYRINR 53.64 1.45 106.37 16.48Aneurinbacillus ASDDAAGLAIS spp. XH2 SEQ ID NO: 300 P spp.Flg22-B6GKLSSGLRING 103.61 37.59 132.95 54.72 Paenibacillus ASDDAAGLAISspp. strain HW567 SEQ ID NO: 293 L spp.Flg22-L2 EKLSSGLRINR 113.04 28.89138.86 53.66 Lysinibacillus spp. AGDDAAGLAIS SEQ ID NO: 574 FLG22-Syn01polypeptide 148.52 6.30 132.35 53.99 +B2+B4 combinations as +B6+L2described above FLG22B2 polypeptide 128.31 0.65 139.74 55.00 +B4combinations as +B6+L2 described above FLG22-Syn01 polypeptide 122.8129.81 124.51 67.31 +B2 combinations as +B6+L2 described aboveFLG22-Syn01 polypeptide 119.17 8.02 100.97 25.95 +B4 combinations as+B6+L2 described above FLG22-Syn01 polypeptide 124.67 8.69 103.45 34.03+B6+L2 combinations as described above FLG22-Syn01 polypeptide 143.027.08 120.67 24.76 +B2+B4 combinations as described above

Example 18: ROS Activity Assay with Cellobiose Additive—Corn and Soybean

Cellobiose is a glucose disaccharide and a building block for cellulosepolymer. Chemically, it is glucose-beta-1-4-glucose, a reducing sugarthat consists of two β-glucose molecules linked by a 13 (1-4) bond.Cellobiose is obtained by the breakdown of cellulose or lichenin andyields glucose upon hydrolysis. Treatments using Bt.4Q7Flg22 werecompared with and without cellobiose in ROS activity assays to determineif cellobiose can act an elicitor to increase ROS production inreactions containing Flg22 polypeptide. The specific treatmentsconducted using ROS assays with corn (FIG. 6, panel A) and soybean (FIG.6, panel B) leaf assays were: Bt.4Q7Flg22 at 25 nM; Bt.4Q7Flg22 at 25nM+cellobiose at 100 μM; Bt.4Q7Flg22 at 25 nM+cellobiose at 1 mM; 100 mMsodium phosphate buffer control; and cellobiose alone (100 μM).

Corn and soybean leaf tissues were harvested from plants as previouslydescribed in Example 15. Flg22 bioactive priming polypeptide stocks wereprepared in either sterile, deionized water or 100 mM sodium phosphate(pH 7.8-8.0) buffer with 0.1% Tween-20. After 18-24 hours, the water wasremoved from each well of the 96-well plate. Samples were treated with a100 μL solution containing Bt.4Q7Flg22 (SEQ ID NO: 226, 25 nM),cellobiose (100 μM or 1 mM), 34 μg/mL luminol, and 20 μg/mL horseradishperoxidase. Recognition of the Flg22 polypeptide by the plant tissueresulted in activation of immune signaling and the production ofapoplastic reactive oxygen species (ROS). In the presence of ROS (H₂O₂),horseradish peroxidase catalyzed the oxidation of luminol and productionof visible light. Relative Light Units (RLUs) were recorded with aGLOMAX 96 microplate luminometer (Promega Corporation) using 0.5 sintegration; 2.6 min intervals over a time course of 40 minutes.

For data analysis, the average RLU per treatment (n=6-16 samples,+/−standard error of the means) was graphed over the time course (FIG.6). Significant outliers beyond the interquartile range were excludedfrom analysis.

The average RLU across the experiment for each treatment is graphed inFIG. 6, panel A (corn) and panel B (soybeans). While ROS production wasobserved in both plant tissue in treatments only containing the Flg22polypeptide (white circles), the addition of cellobiose at 1 mM resultedin significant ROS activity in both plant tissues (black circles).Addition of cellobiose at lower concentrations (100 uM) did not alterROS activity compared to Bt.4Q7Flg22 alone (comparison of white and greycircles in FIG. 6, panel A) and did not lead to any ROS production insoybean (grey circles in FIG. 6, panel B). Notably the combination ofBt.4Q7Flg22 at 25 nM and cellobiose at 1 mM resulted in more ROSproduction in soybean (ROS peak at approximately 25,000 RLU) as comparedto corn leaves (ROS peak at approximately 75,000 RLU).

Example 20: Application of Phytosulfokine (PSKα) to Increase Yield—Cornand Soybean

The effect of Phytosulfokine alpha (PSKα), a sulfonated bioactivepriming polypeptide derived from Arabidopsis thaliana, on corn andsoybean yield was tested. Corn and soybeans were cultivated in the fieldas described in Example 1 and 3. Arabidopsis thaliana PSKα (SEQ ID NO:598) was applied at a final concentration of 1 μM in foliar spray with asurfactant and provided using a uniform application to the above groundplant parts of corn (hybrid 5140RR) and soybean (hybrid 375 NR). At.PSKαformulations were applied at the V5-V8 stage of development in corn andthe V1-V4 stage of development in soybean. Corn and soybean plantstreated with At.PSKα were compared to non-treated control plants(water). Treated plants were randomized at one location in fourreplicate blocks for comparisons to the controls. Yield was reported inBushels per acre (Bu/Ac).

Table 36 depicts how foliar application of At.PSKα resulted in yieldincreases in both corn and soybean yield trials. Both corn and soybeanhad positive yield increases in the field with foliar formulationscontaining At.PSKα applied at the V5-V8 stage of development in corn andthe V1-V4 stage of development in soybean. On average, corn had a +3Bu/Ac (188.3 kg/Ha) increase in overall yield in the field and soybeanhad a +0.8 Bu/Ac (53.8 kg/Ha) yield increase.

TABLE 36 Foliar application of At.PSKα to corn and soybean result inyield increases (Bu/Ac) Bu/Ac Foliar Bu/Ac Foliar Corn Soybean 3.0 0.8

Example 21: Foliar Application of Phytosulfokine Alpha (PSKα) toIncrease Yield—Soybean

A method is provided wherein applying At.PSKα as a foliar application toactively growing soybean plants provides a yield advantage inenvironments with heat and drought stress.

Soybean plants were grown as described in Example 6. The At.PSKαpolypeptide (SEQ ID NO: 598) was applied as a foliar spray to the plantsat the V1-V4 stage. Soybean plants treated with foliar applications ofAt.PSKα and control plants treated with water and surfactant alone werethen grown in conditions described in Examples 7-9 that produced anon-stress and stress (heat and water deficit) environments. Table 37describes the percentage change or increase in height reported forsoybean plants treated with At.PSKα as a foliar spray at the V1-V4growth stage. At.PSKα application resulted in a +3.5% increase in heightin the non-stress environment and a +4.3% increase in height in stressenvironments reported in Bushels per acre (Bu/Ac) as compared to thenon-treated control soybean plants.

TABLE 37 Yield increases in soybean treated with a foliar application ofAt.PSKα and grown in non-stress and stress environments Percentage (%)change in Percentage (%) change in Height in Non-Stress Height in StressEnvironment Soybean with Environment Soybean with At.PSKα over controlAt.PSKα over control 3.5% 4.3%

Example 22: Application of RHPP to Alter Plant Architecture—Corn

Root hair promoting polypeptide (RHPP, SEQ ID NO: 600) originallyderived for soybean (Glycine max) is provided as a foliar application toproduce beneficial phenotypes in corn.

Native and retro inverso RHPP (SEQ ID NOs 600-601) will be applied tocorn plants at the V5-V8 stages. Retro inverso RHPP may be modified withC-terminal amidation prior to application. Treatment with RHPP in thisway is expected to result in a distinct leaf architecture phenotype withan upright leaf orientation and more erect leaves. The increase in leafangle has impactful advantages for use in agriculture in this area. Thisis particularly relevant with higher planting densities used to maximizeyield in a field environment. Foliar applications of the RHPPpolypeptide in maize (corn) is useful for changing the leaf angle thuscontributing to a smaller leaf angle which results in an upright leaforientation. This phenotype can be beneficial for increasing the leafarea index, reducing maize shade syndrome, and improving photosyntheticefficiency. In addition, providing RHPP as a foliar formulation tomaximize canopy development and total light penetrance is key toincreasing vegetative growth of the plants prior to the initiation ofthe grain filling stage.

Example 23: Application of RHPP to Increase Root Biomass and YieldParameter-Soybean

Effective nodulation of soybean roots result in higher yields and higherquality seed production, protein and oil per seed or acre basis. Thiscould be due to increased nitrogen fixation since nodulale formationincreases nitrogen fixation. To determine whether root hair promotingbioactive priming polypeptide, RHPP (SEQ ID NO: 600) could modulate rootbiomass and nodulation and thereby improve nitrogen fixation, soybeanplants (hybrid Morsoy 38×52 and Beck's hybrid 297R4) were treated withfoliar application of RHPP (300 nM) at the R1-R2 stage of development.

Increased Plant Biomass and Nodulation

RHPP bioactive priming polypeptide (SEQ ID NO: 600, originally derivedfrom Glycine max) was applied as foliar treatment to 4-week-old hybridsoybean (Morsoy variety) with 0.1% (v/v) non-ionic surfactant (ALLIGARESURFACE™) using a spray bottle and delivering approximately 1.25ml/plant. The experiment was conducted using a total of 8 plants pertrial per treatment group. The pots were kept in an artificial lightedgrowth room receiving a light level of approximately 300 μmol m⁻² s⁻¹for a 16/8 light/day cycle and a 21° C. day/15° C. night temperaturerange. Growth parameters of nodule counts, root biomass and totalbiomass per plant were measured at 15 days post the foliar applicationand compared between the foliar treatments consisting of ALLIGARESURFACE surfactant (0. 1% v/v) as a control and the RHPP polypeptide(300 nM) containing the ALLIGARE SURFACE surfactant (0. 1% v/v). Averagegrowth parameters as described were normalized to the control plantsthat received the surfactant alone treatment (Table 38).

Nodulation counts on the roots of each plant treated with a foliarapplication of RHPP were compared to the number of nodules on thecontrol plants treated with 0.1% (v/v) surfactant alone. RHPP treatmentresulted in approximately two times the number of nodules on the rootsof each soybean plant compared to control (surfactant) treatment.Soybean plants receiving the foliar application of the RHPP polypeptidealso exhibited an increase in root biomass and total overall plantbiomass which when normalized to the control resulted in an increase ofmore than 20% in root biomass and 8% in total biomass.

TABLE 38 Increases in plant biomass and nodulation in soybean (Morsoyvariety) after foliar application with RHPP bioactive primingpolypeptide (n = 8 replicate plants) RHPP (300 Control nM + RHPPtreatment (surfactant 0.1% 0.1% v/v normalized as a Growth v/v ALLIGAREALLIGARE percentage of the Parameters SURFACE) SURFACE) surfactantcontrol Average nodule 8.88 15.13 170.42% count per plant Root biomass(g) 1.76 2.13 120.57% Total biomass (g) 42.64 46.24 108.44%

Increased Plant Growth

RHPP bioactive priming polypeptide (SEQ ID NO: 600) was also applied asfoliar treatment to R1 stage hybrid soybean (Beck's 297R4) with 0.1%(v/v) non-ionic surfactant (ALLIGARE SURFACE) using a spray bottledelivering approximately 1.2 ml/plant. This experiment was performed tolook at the effects of RHPP on plant growth and was conducted using atotal of 18 plants per treatment group. The pots were kept in anartificial lighted growth room receiving a light level of approximately300 μmol m⁻² s⁻¹ for a 18/6 light/day cycle and a 21° C. day/15° C.night temperature range. R1 stage soybean plants were treated withnothing (non-treated control), ALLIGARE SURFACE surfactant applied at aconcentration of 0. 1% (v/v) or the RHPP polypeptide (300 nM) applied incombination with ALLIGARE SURFACE surfactant (0. 1% v/v). Height foreach plant was recorded at the time of spray and again at 16 days postfoliar application and average growth parameters were compared betweenfoliar treatments (Table 39).

Soybean plants that received the foliar application of RHPP polypeptide(300 nM+0.1% ALLIGARE SURFACE) had increased plant growth (plant height)and an increased change in plant height as compared to the plants thatreceived the surfactant alone and non-treated control (Table 39).

TABLE 39 Increases in plant growth in soybean (Beck's 297R4) with foliarapplication with RHPP bioactive priming polypeptide (n = 18 replicateplants) Control (surfactant RHPP Non- 0.1% (300 nM + 0.1% treatedALLIGARE ALLIGARE Growth Parameters Control SURFACE) SURFACE) Height(cm) 19.3 19.4 19.9 Change in height (cm) 4.0 3.7 4.6

Example 24: Application of RHPP in Combination with a Fertilizer—Soybean

The Gm.RHPP bioactive priming polypeptide (SEQ ID NO: 600) was appliedas a foliar application with a liquid foliar fertilizer, N-RAGE MAX(21-1-3 N—P-K), to two soybean varieties (AG3536 and AG3832). Foliarapplication of RHPP was applied at 1 Fl. oz/Ac or 73.1 mL/Ha (300 nMconcentration) with the recommended use rate of the fertilizer forsoybeans (1 to 2 gal/Ac (9.4 to 18.8 L/Ha), or equal to Nitrogen 2.16lbs/gal (0.29 kg/L); Phosphate P₂O₅ 0.10 lbs/gal and soluble potash (K₂O0.31 lbs/gal or 0.4 kg/L). Foliar application of the combination RHPP,fertilizer treatment was provided to two soybean varieties (AG3536 andAG3832) at the R2 stage (recommended stages R1 to R6) of development in5 locations across the US Midwest (IA, IL, IN). Foliar application ofGm.RHPP with the N-RAGE MAX provided a yield advantage of 1.9 Bu/Ac(127.8 kg/Ha) compared to the control treatment and on average a 1.4Bu/Ac (94.2 kg/Ha) increase compared to those plants that received thefertilizer alone treatment for variety 1 (AG3536) (Table 40).

TABLE 40 Application of RHPP plus a fertilizer Average Average Bu/AcAverage Average Total Change Total Total Yield compared ApplicationYield Yield Bu/Ac to Control Treatment Use Rate Bu/Ac Bu/Ac Variety 1Variety 1 Soybean Fl. oz/Ac Variety 1 Variety 2 and 2 and 2 Control —62.50 62.16 62.33 — N-rage 128 63.04 60.29 61.66 −0.67 Max RHPP 4.065.42 61.04 63.23 +0.9 RHPP + 4.0 64.40 60.96 62.68 +0.35 N-RAGE 128 MAX

Example 25: Application of RHPP Bioactive Priming Polypeptides toTomatoes—Increased Yield

Foliar application treatments of Gm.RHPP (SEQ ID NO: 600) was applied asan exogenous spray at the pre-bloom stage and used to increase yield intomatoes. Two tomato hybrids (JetSetter and Better Big Boy) were plantedin small scale plots as described in Example 12. Foliar treatment ofGm.RHPP was applied at an application use rate of 1 Fl. oz/Ac (73.1mL/Ha) and 20 Fl. oz/Ac(1461.5 mL/Ha) to the two hybrids, JetSetter(Trial 1) and Better Big Boy (Trial 2), at early bloom (first flower)stage. Replicated trials were conducted at the US Midwest (Missouri) inJuly. The foliar treatment of Gm.RHPP on tomato plants was compared tothe control (water applied at same use rate). Effects of the foliartreatments on increasing yield in tomatoes were determined and reportedas normalized to the water control treatment and reported as the averagepercentage change in yield over the average control yield in Table 41.

The average yield represented as a percent change over the controlplants was reported separately for the two trials and as the average forthe two tomato hybrids. Foliar application using Gm.RHPP resulted in anincrease in tomato fruits for each of the two trials when applied at ause rate of 1 Fl. oz/Ac (73.1 mL/Ha). Application of Gm.RHPP resulted inan average increase in tomato yield of +52% over the control plants forthe two hybrids with individual average increases of +93% for theJetsetter hybrid and +10% for the Better Big Boy compared to the controlplants.

TABLE 41 Foliar treatment of RHPP to increase yield in different hybridsof tomato Trial 1: Trial 2: Percent Percent Change in Change in AverageTrials Yield over Yield over 1 & 2 Avg. Control; Avg. Control; PercentChange Hybrid: Hybrid: Better Yield over Avg. Foliar Treatment JetsetterBig Boy Control Gm.RHPP +93% +10% +52% (1 Fl. oz/Ac)

Example 26: Application of RHPP to Peppers—Increased Yield

Foliar treatment of Gm.RHPP (SEQ ID NO: 600) was applied as an exogenousspray at the first-bloom stage to increase yield in two peppervarieties. Foliar treatment of Gm.RHPP was applied using small scaleplots designed to simulate commercial growing conditions for peppers(Capsicum) as described in Example 13. Foliar applications with theGm.RHPP bioactive priming polypeptide were applied at the first flowerstage, on two varieties of pepper, Red Knight (RK) and Hungarian Hot Wax(HHW). The foliar Gm.RHPP treatments were applied using an applicationuse rate of 1 Fl. oz/Ac (73.1 mL/Ha) on the RK and HHW pepper plants andcompared to the control (water applied at same use rate). Effects of thefoliar applications on pepper yield were determined for two separateharvests using a once over harvest approach and normalized to the yieldof the control plants. The average percentage change in yield over theyield for the control plants is reported in Table 42, as the percentchange per total weight (lbs/Ac) of peppers harvested. Average percentchange in yield is reported for the 2 replicate harvests (trials) forthe RK and HHW pepper varieties and then as a combined average for bothvarieties.

TABLE 42 Foliar treatment of RHPP to increase yield in differentvarieties of pepper Avg. Percent Combined Avg. Percent Change Yield Avg.Percent Change Yield Total Weight Change Yield Total Weight (lbs/Ac)Total Number (lbs/Ac) Hungarian (lbs/Ac) Foliar Treatment Red Knight HotWax RK and HHW Gm.RHPP +87% +46% +67% 1 Fl. oz/Ac

Percent average yield for RK and HHW peppers that received the Gm.RHPPapplied at the use rate of 1 Fl. oz/Ac (73.1 mL/Ha) was increased by 87%for RK and 46% for HHW peppers compared to the control plants. Thecombined average for both pepper varieties was reported as an average67% increase for the percent change in yield in the foliar Gm.RHPPtreated peppers over the non-treated (water) control pepper plants(Table 42).

Example 27: Application of Harpin-like and ALPSKα Polypeptides to Corn

Harpins can provide functional benefits when applied both exogenously,for example as a foliar spray to the plant surface, or providedapoplastically (the space outside of the plant cell membrane) orendogenously (inside a plant cell/plant cell membrane). Synthetic harpinbioactive priming polypeptide, HpaG-like (Xanthomonas spp., SEQ ID NO:587) was applied exogenously to the surface of corn plants at the V2-V3stage of development. Additionally, the effect of exogenous applicationof Phytosulfokine alpha (PSKα), a sulfonated bioactive primingpolypeptide derived from Arabidopsis thaliana, on corn growth wastested.

Corn (Beck's hybrid 5828 YH) plants were grown in an environmentallycontrolled growth room. Corn seed was planted directly into 39.7 cm³pots containing Timberline top soil at a depth of 2.54 cm, with 2 seedsper pot. After planting, 50 mL of room temperature water was added toeach pot to allow for germination. The pots were kept in an artificiallighted growth room receiving approximately 300 μmol m⁻² s⁻¹ (lightphotons) for a 16/8 light/day cycle and a 21° C. day/15° C. nighttemperature range. Plants received the same watering and fertilizerregimes.

Plant height (cm) was measured at 3 weeks after emergence. Bioactivepriming polypeptides for HpaG-like (SEQ ID NO: 587), provided as asynthetic 23 amino acid polypeptide, and At.PSKα (SEQ ID NO: 598) werethen applied to the corn plants as a foliar spray at finalconcentrations of 1 μM for HpaG-like and 100 mM for PSKα bioactivepriming polypeptides. Control plants were treated with surfactant (0.01%v/v) alone. A week after the spray treatments were applied, the plantswere subdivided into 2 groupings where one group remained in the samestandard growth environment described above and the other group wastransferred to an environment that provided heat and water deficitstress. For the heat and water deficit treatments, the growth roomenvironment (with the exception of temperature and watering/fertilizercycles) remained similar to the standard growth environment). Heatstress was applied using heat mats to raise the temperature in theenvironment from 21° C. to 27° C. During the period of heat stress, theplants were left unwatered to simulate a water deficit stress. Change inplant height (cm) was measured at 5 weeks and reported as normalized toor as a percentage of the height of the control (water) plants.Measurements are reported as the combined average of two trials with 9replicate plants per trial (Table 43) and are presented as a percentageof growth over control corn plants that received water plus surfactant(0.01% v/v) standardized to measure 100% (Table 43).

TABLE 43 Changes in Plant height of corn plants treated with X. spp.HpaG-like and At. PSKα Plant Height Plant Height Normalized NormalizedHeight Height as a as a Height (cm) (cm) percentage percentage (cm) andafter non- after of control of control Treatment (STDEV) stress stressheight height Corn 3 weeks 5 weeks 5 weeks Non-stress Stress X, spp.47.23 64.10 45.50 107.4% 89.4% HpaG-like (6.11) (5.53) (4.37) (1 μM)At.PSKα 49.36 58.62 54.88 98.2% 107.8% (100 nM) (8.00) (4.84) (2.79)

Foliar application using the HpaG-like polypeptide showed an improvedgrowth phenotype in normal environments, but not stressed environments,when compared to the control plants, while foliar application of PSKαexhibited an improved growth phenotype when grown under conditions ofheat and water deficit stress but not in the non-stressed environment.

In a separate set of replicated trials, similar changes in growth ratesresulted from the foliar applications of HpaG-like (SEQ ID NO: 587) andPSKα (SEQ ID NO: 598). Table 44 shows the percentage change in plantgrowth for corn receiving X. spp. HpaG-like polypeptide (1 μM finalconcentration) and At.PSKα (100 nM final concentration) applied asfoliar treatments and measured by changes in plant height compared tocontrol (water plus 0.01% v/v surfactant) plants grown in optimal(non-stress) and in stress environments. This suggests that the combinedfoliar application or sequential applications of PSKα with HpaG-likebioactive priming polypeptides may be useful for enhancing growth ofplants growth under standard (non-stress or optimal growth) environmentsor of plants exposed to abiotic stress (for example, heat, and waterdeficit stress).

TABLE 44 Foliar application treatments using the Xspp HpaG-like and theAt.PSKα polypeptides on corn grown under non-stress and stressconditions Plant Height (cm) Plant Height (cm) Percentage ChangePercentage Change Compared to Control Compared to Control (0.01%surfactant) (0.01% surfactant) Treatment Non-Stress Stress Xspp.HpaG-like (1 μM) +5.0% −6.1% At. PSKα (100 nM) −11.8% +6.1%

Example: 28 Combination of Bt.4Q7Flg22 or Ec.Flg22 with RHPP

The bioactive priming polypeptides, Bt.4Q7Flg22 and Ec.Flg22, werecombined with RHPP and accessed for yield benefits in soybean. Thecombination of either Bt.4Q7Flg22 (SEQ ID NO: 226) or Ec.Flg22 (SEQ IDNO: 526) and RHPP (SEQ ID NO: 600) were foliar applied to two varietiesof soybean (AG2836, Variety 1; AG3536, Variety 2) in 7 locations acrossthe US Midwest (IA, IL and IA).

Foliar application using Bt.4Q7 Flg22 bioactive priming polypeptide (SEQID NO: 226; FIG. 4, panel A) and Ec.Flg22 (SEQ ID NO: 526; FIG. 4, panelB) and RHPP (SEQ ID NO: 600) were applied individually to soybean plants(commercial hybrid Beck'S 294 NR) at the R2 stage of development usingvarying use rates of 0.33, 4.0, 8.0, and 16.0 Fl. oz/Ac or (24.1 mL/Ha,292.3 mL/Ha, 584.6 mL/Ha, 1169.2 mL/Ha). Average yield (harvested inSeptember) in bushels per acre (Bu/Ac) is reported for soybean grown in7 separate locations and reported individually for both soybeanvarieties and as a combined average yield (Table 45). Soybean yield(Bu/Ac) is also reported as the change in yield (Bu/Ac) normalized tothe control soybean plants for both varieties.

TABLE 45 Flg polypeptides and RHPP polypeptides increase yield insoybean Average Average Bu/Ac Average Average Total Increase Total TotalYield compared to Application Yield Yield Bu/Ac Control Treatment UseRate Bu/Ac Bu/Ac Variety 1 Variety 1 Soybean Fl. oz/Ac Variety 1 Variety2 and 2 and 2 Control — 59.53 61.61 60.57 — Bt.4Q7Flg22 0.33 60.33 61.6161.02 +0.45 Bt.4Q7Flg22 4.0 57.61 64.19 60.90 +0.33 Bt.4Q7Flg22 8.059.05 63.86 61.45 +0.88 Ec.Flg22 0.33 58.62 63.58 61.10 +0.53 Ec.Flg224.0 58.02 63.91 60.74 +0.17 Ec.Flg22 8.0 58.27 64.35 61.31 +0.74 Gm.RHPP0.33 59.15 62.44 60.92 +0.35 Gm RHPP 4.0 58.61 66.35 61.83 +1.26 Gm RHPP8.0 59.47 62.46 61.08 +0.51 Bt.4Q7Flg22 + 4.0 61.14 64.88 63.18 +2.61Gm.RHPP 4.0 Ec.Flg22 + 4.0 59.56 62.46 61.08 +0.51 Gm.RHPP 16

Soybean variety AG3536 (Variety 2) consistently outperformed AG3536(Variety 1) for yield Bu/Ac in all 7 locations across the US Midwest.Foliar applications with the Bt.4Q7Flg22, Ec.Flg22 and RHPP appliedindividually at the 3 different use rates (0.33, 4.0 and 8.0 Fl. oz/Ac)or (24.1 mL/Ha, 292.3 mL/Ha, 584.6 mL/Ha) all resulted in a yieldadvantage over the non-treated control plants. The RHPP applied foliarlyusing a 4.0 Fl. oz/Ac (292.3 mL/Ha) use rate resulted in the largestyield increase of +1.26 Bu/Ac (84.7 kg/Ha) over the control plantscompared to the other bioactive priming polypeptides applied separately.However, the combination of Bt.4Q7Flg22 with RHPP provided an additionalyield advantage resulting in a +2.61 Bu/Ac (175.5 kg/Ha) over thenon-treated soybean control plants. This increase in yield seen fromsoybean plants treated with foliar applications of Bt.4Q7Flg22 combinedwith RHPP illustrates a synergistic effect achieved by combining thebioactive priming polypeptides where the increase in yield of thecombination was greater than the sum of the two polypeptides appliedseparately.

Example 29: Use of Agrobacterium tumefaciens to Test Effectiveness ofThionins in Treating HLB Disease

Agrobacterium tumefaciens strain GV3101 was inoculated into Luria brothmedium (LB) and grown for 20 hours. Initially the optical density (OD)of the culture was measured at a wavelength of 600 nm using aspectrophotometer and normalized to a low starting density. The cultureswere then divided equally and treated with similar proportions ofthionins that are representative of mixtures used to treat citrus trees.The ratios of Cs.thionin (SEQ ID NO: 651), As.thionin (SEQ ID NO: 652)and Mt.thionin (SEQ ID NO: 653) used were 10.0%, 2.0%, 0.40%, 0.08%, and0.02% and were prepared to match the 20 mL total volume of filtrate ofeach of the thionin mixtures that is used as a treatment per tree. Eachthionin mixture was also compared to control mixtures containing only:filtrate, minimal media (LB), or a tetracycline (Tet) antibiotic (10μg/mL per culture). Each bar represents a combined OD measure of 3replicates. After incubation with the thionin and antibiotic mixtures,the optical density (OD 600) was measured again to determine if growthof the Agrobacterium cultures was reduced or inhibited.

As is shown in FIG. 8, the Cs.thionin, As.thionin and Mt.thionintreatments all showed a dose dependent response and decreased growth ofthe Agrobacterium cultures compared to the filtrate, minimal media (LB)or antibiotic (Tet) controls.

Example 30: Treatment of Candidatus liberibacter asiaticus Infectionwith Thionins

Use of thionins to treat Candidatus liberibacter asiaticus infectionwill be tested in citrus trees from an orchard located in centralFlorida (Okeechobee county). Treatment of a total of 26 trees will useformulation mixtures of thionin (SEQ ID NO: 620; 621 and 622) eitherwith or without a phloem localization sequence (SEQ ID NO: 611) totarget the thionins specifically to the phloem where Candidatusliberibacter asiaticus reside. Inoculation of Valencia orange (Citrussinensis) trees with these formulations of thionins and mixtures thereofwill be conducted using a low-pressure injection device, BRANDT ENTREE.Four total thionin treatments including water as a negative control andoxytetracycline as a positive control will be applied to 5 year-oldtrees. The citrus trees will be randomized into treatment blocks forcontrol (non-treated), thionin treated and positive control antibiotic(oxytetracycline) treated tree plots. Thionins fused to a phloemtargeting sequence will be expressed in a pBC vector, and thionincontaining filtrate will be collected from the expressed cells. A totalvolume of 20 mL containing a mixture of thionins: Cs.thionin (SEQ ID NO:651), As.thionin (SEQ ID NO: 652) and Ms.thionin (SEQ ID NO: 653) willbe provided as 20 mL total volume of filtrate. The thionin treatedcitrus trees will be compared to the non-treated (control) trees andtrees that received a separate positive control of an antibiotic,oxytetracyline, applied with a concentration of 2 grams/tree. Levels ofinfection of trees with Candidatus liberibacter asiaticus will beconfirmed by qPCR detection or amplification using 16S rRNA genespecific primers and nested primers to detect the HLB disease [Sequence5′>>3′:(forward) HLB as TCGAGCGCGTATGCAATACG; (reverse) HLBrGCGTTATCCCGTAGAAAAAGGTAG; HLBpc (probe) AGACGGFTGAGTAACGCG labeled withfluorescein reporter dye].

Plants will be treated in March and leaf samples will be collected onemonth later in April. Average bacterial counts for Candidatusliberibacter asiaticus will be assessed along with visual symptomologyranking scores for leaf blotch mottling or signs of yellowing of leavesand stems.

Fruit size, shape and level of fruit development or maturity will becollected for 20 representative fruits per tree. Longitudinal length(major diameter, cm) and width (minor diameter, cm, the average of thelargest and smallest widths if the fruit is not symmetrical). Fruitshape will be measured by the ratio of width to length. Total fruitweight will be obtained and divided by the total number of fruits (20)to provide an average fruit weight (grams). Total fruit weight will becollected and represented in kg/tree.

Acid-corrected ° Brix (° Brix_(c)) values of juice obtained from thejuiced (squeezed) grapefruit and orange fruit will be obtained per treefollowing the USDA minimum standards for ° Brix_(c) laboratoryanalytical methods. Percent acid (%, w/v) will also be measured. The °Brix reading on a refractometer for a juice to be reconstituted equalsthe value of the desired acid-corrected ° Brix subtracted of the acidcontribution and temperature effect. The total titratable acidity (%acid) of the reconstituted juice will also be calculated based on thereconstituted ° Brix and Brix/Acid ratio and adjusted using an acidcorrection and temperature correction factors (JBT FoodTech LaboratoryManual, “Procedures for Analysis of Citrus Products, Sixth Edition).

Bacterial cell counts will be calculated using real time fluorescentPCR, quantitative polymerase chain reaction (qPCR) techniques to detectonly live bacterial and subtract out background DNA including naked DNAor DNA from dead cells (Davis and Brlansky, “Quantification of liveCandidatus liberibacter asiasticus” populations using real-time PCR andpropidium monoazide”, Plant Disease 97: 1158-1167, 2013). Colony countsspecific for Candidatus liberibacter asiasticus (CLas) cells will bemeasured in the leaves collected from the thionin treated, non-treated(control) and positive control (antibiotic) treated trees. Calculationsfor live bacterial titers will be obtained from the DNA yield obtainedby qPCR, fit into a regression equation to correlate target copy numberto total bacterial counts and represented on a log scale of live cellsper gram tissue. Comparisons of titers from treated and non-treatedtrees will be matched with the degree of disease severity or diseasesymptoms, such as the classic blotchy mottling on the leaves, deformedor lopsided fruit and greening fruit, etc. for both the red grapefruitand Valencia orange trees.

Example 31: Use of Retro-Inverso Flg Bioactive Priming Polypeptides toTreat and Reduce Citrus Greening

Combinations of flagellin-associated polypeptides paired with theirretro-inverso counterparts can be used to treat and reduce the greeningeffect on citrus that results in Asian citrus greening or Huanglongbingdisease (HLB).

An early symptom of HLB in citrus is the yellowing of leaves on anindividual limb or in one sector of a tree's canopy. Leaves that turnyellow from HLB will show an asymmetrical pattern of blotchy yellowingor mottling of the leaf, with patches of green on one side of the leafand yellow on the other side. As the HLB disease progresses, the fruitsize becomes smaller, and the juice turns bitter. The fruit can remainpartially green and tends to drop prematurely.

The retro-inverso forms of Flg22 can compete with native forms of Flg22for binding to the FLS-associated receptor(s) at the plant surface andthus inhibit/delay the symptom formation of greening associated with HLBdisease. Using native Flg22 and RI combinations will assist with a finetuned immune response to reduce and even eliminate the disease-causingbacteria, Candidatus liberibacter asiaticus and thus prevent acutesymptom development, such as leaf yellowing and citrus fruit greening.

Treatment combinations of Flg polypeptides with their retro-inverso (RI)forms will be used to minimize the effect of HLB infection on citrusfruit greening. Thirty-four commercial grapefruit, Citrus paradiseMacfad., and six sweet orange, Citrus sinensis (L.) trees, with orwithout symptoms of HLB disease, will be treated using flagellinbioactive priming polypeptide combinations described in Table 46, below,using a low pressure injection device called BRANDT enTREE to distributethe Flg polypeptides into the interior of the tree.

TABLE 46 Combinations of Flg22 native and retro-inverso Flg22 bioactivepriming polypeptides Treatments SEQ ID NO: Concentration nM Bt.4Q7Flg22226 50 nM Bt.4Q7Flg22 226 100 mM RI Bt.4Q7Flg22 376 50 nM RI Bt.4Q7Flg22376 100 mM Bt.4Q7Flg22 + RI 226 & 376 50 nM Bt.4Q7Flg22 Bt.4Q7Flg22 + RI226 & 376 100 mM Bt.4Q7Flg22 Ec.Flg22 526 50 nM Ec.Flg22 526 100 mM RIEc.Flg22 527 50 nM RI Ec.Flg22 527 100 mM Ec. Flg22 + RI Ec.Flg22 526 &527 50 nM Ec. Flg22 + RI Ec.Flg22 526 & 527 100 mM

Leaf tissue samples from these treated grapefruit and sweet orange treeswill be analyzed using the ROS assay as described in Example 15.Sampling will be conducted in orchard groves from March to August incentral Florida. The sample of citrus orchards will be assumed to berepresentative of the state. The orchard sampled will have a minimumacreage of 2 hectares (range of 2-24 Ha and an average of 5.2 Ha).Selected orchard citrus trees will be randomly selected with thenon-treated control trees nested in each randomized plot. Leaf tissuesfrom the grapefruit and orange trees will be collected from trees ofapproximately the same age. Leaves will be sampled at similar locationson the trees and only from trees that had a new flush of growth at thetime of sampling. In the orchards selected for sampling, similarcultural practices will be maintained and include flood irrigation andweed management with herbicides. However, the selected orchards will notreceive any pesticide application for a minimum of 30 days before leafsampling for the ROS assays. Two replicate trials of 10 grapefruit treesexhibiting symptomology of HLB disease will be randomly sampled perorchard and compared to 14 grapefruit trees (non-infected control)sampled that do not exhibit any symptoms. Similar leaf sampling will beperformed in sweet orange (four infected samples compared to 2uninfected controls). Trees will be selected to be representative of thewhole orchard. A nested analysis of variance (ANOVA) will be performedto determine the statistical significance of any differences in ROSactivities observed from treatment of the control and infected HLBcitrus leaf samples.

Example 32: Foliar Application of the Flg22 Polypeptide ReducesCercospora Leaf Blight Disease of Soybean

Foliar application of the Bt.4Q7Flg22 bioactive priming polypeptide (SEQID NO: 226) derived from Bacillus thuringiensis and Bacilluspseudomycoides expressing Bt.4Q7Flg22 (H1) were applied to soybeanplants (commercial hybrid Beck's 294 NR) at the V3 stage of developmentthat were grown at 3 separate US Midwestern locations that were known topreviously have Cercospora infection in the fields.

A Cercospora leaf blight rating scale (percentage of leaf area affected)was used to rate disease severity in all field experiments. Thepercentage of leaf area affected was calculated using a visual key basedon the ASSESS image analysis for plant disease quantification (ChagasFerreira da Silva, LSU Master's Theses, 2014). Symptom ranking as apercentage was done for the uppermost trifoliate leaves

The results are described in Table 47. Visually, soybean plants thatreceived the foliar treatments of the Bt.4Q7Flg22 bioactive primingpolypeptide and Bacillus pseudomycoides expressing Bt.4Q7Flg22 (H1) hadincreased vigor as compared to the non-treated control plants. Thecontrol plants showed an increase in early symptom development at the 4week observation time point, 30% as compared to 20% with the Bt.4Q7Flg22treatment (0.33 Fl. oz/Ac or 24.1 mL/Ha) and approximately 5% with theBt.4Q7Flg22 treatment (4.0 Fl. oz/Ac or 292.3 mL/Ha). Soybean plantsreceiving the Bacillus pseudomycoides expressing the Bt.4Q7Flg22 (H1)treatment also showed less early symptom development as a result ofCercospora infection than the non-treated control plants, 20% at 4 weeks(0.33 Fl. oz/Ac or 24.1 mL/Ha) and 10% (4.0 Fl. oz/Ac or 292.3 mL/Ha).At 8 weeks post application, the non-treated control plants showed 50%visual symptom damage on the upper foliage of the plant (top 3-4trifoliate leaves). The symptom ranking for plants that received thefoliar treatments of the Bt.4Q7Flg22 polypeptide (0.33 Fl. oz/Ac or 24.1mL/Ha) was comparable to the non-treated control plants at 8 weeks postfoliar treatment. However, the soybean plants that received the foliartreatments of Bt.4Q7Flg22 polypeptide (4.0 Fl. oz/Ac) and Bacilluspseudomycoides expressing Bt.4Q7Flg22 (H1) (0.33 Fl. oz/Ac or 24.1mL/Ha) and 4.0 Fl. oz/Ac or 292.3 mL/Ha) showed considerably lessapparent symptoms and damage. Overall the treatment of the Bt.4Q7Flg22polypeptide (4.0 Fl. oz/Ac or 292.3 mL/Ha) was effective at theprevention of early symptom development from Cercospora infection ascompared to the non-treated plants that showed blight and purplecoloration symptoms as well as defoliation. Therefore, foliarapplication of Bt.4Q7Flg22 polypeptide applied at a higher applicationuse rate (eg. 4.0 Fl. oz/Ac or 292.3 mL/Ha)) can provide a means ofmanaging early symptom development and provide healthier more vigoroussoybean plants grown in field locations that have been impacted byCercospora.

TABLE 47 Foliar treatment of soybean plant with Bt.4Q7Flg22 and Bp.Expressing Bt.4Q7Flg22 resulted in disease reduction and symptomdevelopment of Cercospora on soybean Percent of Disease Application Areacovering Disease Area, Treatment- Use Rate plant, 4 weeks 8 weeks PostSoybean Fl. oz/Ac Post Application Application Control — 30% 50%Bt.4Q7Flg22 0.33 Fl. oz/Ac 20% 50% Bt.4Q7Flg22 4.0 Fl. oz/Ac  5% 35% H1Bt.4Q7Flg22 0.33 Fl. oz/Ac 20% 40% H1 Bt.4Q7Flg22 4.0 Fl. oz/Ac 10% 30%

Example 33: Application to Corn—Enhanced Normalized DifferenceVegetation Index (ENDVI) Analysis

Enhanced Normalized Difference Vegetation Index (ENDVI) is an indicatorof live, photosynthetically-active green vegetation and was used tocompare the effectiveness of treatments in field trials using remotesensing technology. In the ENDVI index, values ranging from −1.0 to 0.1,are indicative of unhealthy plants with decreased photosynthesis,whereas values approaching 1 are indicative of lush greenness, highphotosynthetic capacity, and increased biomass. Healthy plants stronglyabsorb visible light from the 400-700 nm spectral wavelength range andreflect the wavelengths in the near-infrared light from 700-1100 nm.ENDVI measurements can correspond to certain vegetative properties, suchas plant biomass or greenness, absorption of light by plant canopies,photosynthetic capacity (e.g., leaf area index, biomass, and chlorophyllconcentration). ENDVI images were collected using a BGNIR camera(Zenmuse X3) attached to a drone (DJI MATRICE 100) specifically createdto capture images and filter different wavelengths of light during thecapture. The camera uses sensors to capture visible and near-infraredbands of the electromagnetic spectrum. Healthy plants with large amountsof vegetation or biomass reflect green (G) and near-infrared (NIR)light, while absorbing both blue (B) and red light. Plants that are lesshealthy or that have less above-ground biomass reflect more visible andless NIR light. ENDVI uses both NIR and G as the reflective channelswhile using B as the absorption channel. The ENDVI formula below addsthe NIR and green channels together for the reflective channel. The bluechannel is multiplied by two to compensate for the NIR and G channelsbeing added together. The ENDVI equation uses the following calculationfor the NIR, G, and B channels to provide a ratio value as a singleoutput.

${ENDVI} = \frac{\left( {{NIR} + {Green}} \right) - \left( {2*{Blue}} \right)}{\left( {{NIR} + {Green}} \right) + \left( {2*{Blue}} \right)}$

Corn seed (DEKALB hybrid DKC 58-89) treated with a seed treatmentcomprising EVERGOL fungicide (7.18% propiconazole, 3.59% penflufen andcombined with 5.74% metalaxyl) and PONCHO/VOTiVO 500 (a mixture of 40.3%clothianidin insecticide and 51.6% Bacillus firmus I-1582, a microbialagent) was planted in the US Midwest (IL). Various foliar treatmentscontaining Bt.4Q7Flg22 and a synthetic version of Bt.4Q7Flg22(Syn01Flg22 as described in Table 48) were applied to corn plants at theV5-V7 stage of development. BGNIR images were collected by drone flight,50 m above the trial plot, three weeks after each foliar treatment andafter the corn canopy had fully closed. Individual BGNIR images wereprocessed using drone display image analysis software to create a singleorthomosaic image of the trial plot that was further analyzed with Fijiimaging software. Within the orthomosaic image, plot regions to identifyindividual foliar treatments in a field and the replicates per eachtreatment were clearly established using GPS coordinates in each fieldtrial. The treatment replicates identified for imaging were consistentin size. For each foliar treatment, three replicates were collected withtwo rows imaged per each replicated plot. Within each replicate, theaverage intensity of light was measured for each of the image channels[blue, green, and near infrared, (visualized as red)] on a scale of0-255, with Intensity 0=0% reflection (black pixel) and Intensity255=100% reflection (white pixel). These average B, G and NIR lightintensities were used to calculate an ENDVI value using the ENDVIalgorithm for plant health (greenness) for each replicated plot. TheENDVI values were then averaged for the three plot replicates asreported in Tables 49 and 50. ENDVI values for the treatmentapplications were compared to the control treatments in each plot.Control treatments consisted of corn plants grown from seed that wastreated with a base seed treatment only and received no foliartreatments. Foliar treatment compositions were as described using theapplication use rates as specified in Table 48.

TABLE 48 Compositions of foliar Flg22 treatments for testing on corn andsoybean Application Use Rate Fluid ounce/acre (Fl. oz/Ac) CompositionFoliar Formulation Milliliters/hectare (mL/Ha) Composition 1Bft.4Q7Flg22 (SEQ ID NO: 226) 16.7 μM 4 Fl. oz/Ac or PROXEL BCpreservative: 330.7 μM (BIT); 292.3 mL/Ha 53.5 μM (CMIT); 26.1 μM (MIT)Composition 2 Bt.4Q7Flg22 (SEQ ID NO: 226) 16.7 μM 4 Fl. oz/Ac or 11.6mM Sodium Phosphate Dibasic 292.3 mL/Ha combined with 4.2 mM Citric AcidMonohydrate pH 5.6 PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT);21.71 μM (MIT) Composition 3 Bt.4Q7Flg22 (SEQ ID NO: 226) 16.7 μM 4 Fl.oz/Ac or 1.67 mM Sodium Phosphate Buffer, pH 292.3 mL/Ha 5.7 PROXEL BCpreservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) Composition 4Bt.4Q7Flg22 (SEQ ID NO: 226) 16.7 μM + 4 Fl. oz/Ac (Flg22) Cellobiose:320 mM 292.3 mL/Ha 1.67 mM Sodium Phosphate Buffer, pH 8 Fl. oz/Ac(Cellobiose) 5.7 584.6 mL/Ha PROXEL BC preservative: 330.7 μM; 50.1 μM(CMIT); 21.71 μM (MIT) Composition 5 Bt.4Q7Flg22 (SEQ ID NO: 226) 16.7μM 48 Fl. oz/Ac or 1.67 mM Sodium Phosphate Buffer, pH 3,507.6 mL/Ha 5.7PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT)Composition 6 Syn01Flg22 (SEQ ID NO: 571) 16.7 μM 4 Fl. oz/Ac or 1.67 mMSodium Phosphate Buffer, pH 292.3 mL/Ha 5.7 PROXEL BC preservative:330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) Composition 7 Syn01Flg22 (SEQID NO: 571) 16.7 μM 0.4 Fl. oz/Ac or 1.67 mM Sodium Phosphate Buffer, pH29.23 mL/Ha 5.7 PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71μM (MIT) Composition 8 Syn01Flg22 (SEQ ID NO: 571) 16.7 μM + 0.4 Fl.oz/Ac (Flg22) Cellobiose: 320 mM 29.23 mL/Ha 1.67 mM Sodium PhosphateBuffer, pH 8 Fl. oz/Ac (Cellobiose) 5.7 584.6 mL/Ha PROXEL BCpreservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) Composition 9At.Flg22-B4 (SEQ ID NO: 300) 4 Fl. oz/Ac or 1.67 mM Sodium PhosphateBuffer, pH 292.3 mL/Ha 5.7 PROXEL BC preservative: 330.7 μM; 50.1 μM(CMIT); 21.71 μM (MIT)

Foliar compositions contained 0.1% (v/v) PROXEL BC preservative, anaqueous dispersion of a blend of 330.7 mM 1,2-benzisothiazolin (BIT),53.5 mM 5-chloro-2-methyl-4-isolthiazolin-3-one (CMIT), and 26.1 mM2-methyl-4-isothiazolin-3-one (MIT). Foliar compositions were applied atthe indicated rates (Fl. oz/Ac or mL/Ha) in a carrier volume of 20gallons/acre water with 0.1% (v/v) Alligare Surface™ non-ionicsurfactant

As shown in Table 49, foliar applications with compositions containingBt.4Q7Flg22 and Syn01Flg 22 applied to corn at the V5-V7 stage ofdevelopment resulted in increased ENDVI measurement ratio values ascompared or normalized to plants that received no foliar treatment (seedtreatment control). The Bt.4Q7Flg22 compositions provided as a foliartreatment in buffered formulations (compositions 2, 3, 4 and 5; sodiumphosphate pH 5.6-5.7) resulted in plants with higher ENDVI ratio valuescompared to the plants that received the Bt.4Q7Flg22 provided as anon-buffered composition (composition 1) applied at 4 Fl. oz/Ac or 292.3mL/Ha. However, ENDVI ration values of Bt.4Q7Flg22 treated plants(compositions 1-5) were all increased relative to the non-treatedcontrol plants. Corn plants that received the foliar treatmentapplication of composition 4 consisting of the Bt4Q7Flg22 polypeptidecombined with cellobiose in a phosphate buffered formulation provided at8 Fl. oz/Ac or 584.6 mL/Ha use rate resulted in a +9% increase in theaverage ENDVI ratio value over the control (seed treatment only) plants.Plants that received foliar applications of composition 1 and 5 whichdiffered in composition only in the respective application use rates (4and 48 Fl. oz/Ac or 292.3 mL/Ha and 3,507.6 mL/Ha) resulted in plantswith similar ENDVI ratio values (+3% and +4%) as compared to the controlplants. Importantly, the higher rate of 48 Fl. oz/Ac or 3,507.6 mL/Haresulted in no detectable phytotoxicity, which would have been observedas a reduced ENDVI value compared to the control (seed treatment only).A synthetic derived variant version of Bt.4Q7Flg22 (Syn01Flg22)compositions 6, 7, and 8 were also provided as a foliar spray to V5-V7corn plants. The Syn01Flg22 polypeptide provided in a phosphate bufferedformulation (composition 6 and composition 7) were compared according tothe application use rates. The Syn01Flg22 polypeptide (composition 6)that was provided to corn plants using a higher application use rate (4Fl. oz/Ac or 292.3 mL/Ha) resulted in a decreased ENDVI ratio value orhad a lesser percentage increase in ENDVI as compared to the Syn01Flg22polypeptide (composition 7) applied to plants using a 0.4 Fl. oz/Ac or29.23 mL/Ha application use rate, a change of 8% between the composition6 and 7 treatments. The Syn01Flg22 (composition 8) had the addition ofcellobiose and similar to composition 7 was provided at an applicationuse rate of 0.4 Fl. oz/Ac or 29.23 mL/Ha. Foliar application ofSyn01Flg22 (composition 7) was compared to Syn01Flg22 combined withcellobiose (320 mM) (composition 8). The Syn01Flg22 composition 7 andcomposition 8 had similar increases in ENDVI measurement ratiosresulting in a +10% increase as compared to control plants or a +8%increase compared to plants that received the Syn01Flg22 (composition 6)provided at the higher 4 Fl.oz/Ac or 292.3 mL per hectare (Ha) use rate.

TABLE 49 ENDVI outputs provided for foliar Flg22 treatments on cornhybrid DKC 52-61 Percentage AVG Change in ENDVI Normalized to TreatmentENDVI (STDEV) Control Plants* Seed Treatment Control 0.253 (0.027) —Bt.4Q7Flg22 Composition 1 0.259 (0.019) +3% Bt.4Q7Flg22 Composition 20.269 (0.021) +6% Bt.4Q7Flg22 Composition 3 0.272 (0.003) +7%Bt.4Q7Flg22 + Cellobiose 0.276 (0.009) +9% Composition 4 Bt.4Q7Flg22Composition 5 0.263 (0.010) +4% Syn01Flg22 Composition 6 0.258 (0.016)+2% Syn01Flg22 Composition 7 0.277 (0.018) +10%  Syn01Flg22 + Cellobiose0.279 (0.015) +10%  Composition 8 *Normalized to seed treatment control:EVERGOL and PONCHO/VOTiVO 500

Corn seed (DEKALB hybrid DKC 52-61) was also treated with RoundupPOWERMAX (active ingredient glyphosate, 48.7% in the form of potassiumsalt) in combination with the Bt.4Q7Flg22 composition 3. RoundupPOWERMAXwas applied using the recommended use rate on the specimen labelof 24 Fl oz/Ac. The Bt.4Q7Flg22 (composition 3) was applied at a rate of4.0 Fl. oz/Ac or 292.3 mL/Ha. Results are shown in Table 50.

TABLE 50 ENDVI with foliar applications of Flg22 polypeptides combinedwith an herbicide on corn (hybrid DKC 52-61) Percentage AVG ChangeTreatment in ENDVI Normalized Formulation Code ENDVI to RoundupApplication Use Rate (STDEV) POWERMAXTreatment Roundup POWERMAX 0.271 —(0.006) Roundup POWERMAX + 0.293 +8% Bt.4Q7Flg22 (0.005) (Composition 3)*Normalized to RoundUp POWERMAX foliar treatment

As shown in Table 50, Roundup POWERMAXapplied to corn at the V5-V7 stageof development as a foliar herbicide combined with the Bt.4Q7Flg22(composition 3) resulted in an increased ENDVI measurement ratio, anincrease of +8% compared to the treatment with the Roundup POWERMAXapplied without the Bt.4Q7Flg22 polypeptide.

Example 34: Application of Bioactive Priming Polypeptides to V4-V7Corn—Increased Yield

Large acre corn trials were planted from corn seed (DEKALB hybrids: DKC52-61, DKC 58-89, and DKC 65-81) coated with a seed treatment comprisingEVERGOL fungicide (7.18% propiconazole, 3.59% penflufen, and 5.74%metalaxyl) with PONCHO/VOTiVO 500 (a mixture of clothianidin insecticideand a microbial agent, Bacillus firmus I-1582). Corn field trials wereplanted in 8 locations throughout the US Midwest (IN, IL, & IA). Fieldseed beds at each location were prepared using conventional orconservation tillage methods for corn plantings. Fertilizer was appliedas recommended by conventional farming practices which remainedconsistent between the US Midwest locations. Herbicides were applied forweed control and supplemented with cultivation when necessary. Four-rowplots, 5.3 meters were planted at all locations. Corn seed was planted3.8 to 5.1 cm deep to ensure normal root development. Corn was plantedat approximately on average of 42,000 plants per acre or 103,782 plantsper hectare with an average row width of 0.8 meters with seed spacing of1.6 to 1.8 seeds per every 30 cm.

Corn plants at approximately the V5 stage of development received foliarapplications using a foliar composition comprising a Bt.4Q7Flg22 (SEQ IDNO: 226) polypeptide and a synthetic version of Bt.4Q7Flg22 which isdescribed as Syn01FfIg22 (SEQ ID NO: 571) polypeptide. The foliarcompositions comprising the Bt.4Q7Flg22 polypeptide and a syntheticversion of Bt.4Q7Flg22 polypeptide were applied to 3 corn hybrids(DEKALB hybrids: hybrid 1: DKC 52-61; hybrid 2: DKC 58-89; hybrid 3: DKC65-81) planted in 8 locations throughout the US Midwest (IN, IL, & IA).Corn plants received foliar treatments using the concentrations andapplication use rates as described in Table 51. Corn yield (Bu/Ac) wascollected and reported as the average yield (Bu/Ac) across the locations(8 locations for hybrid 1, 7 locations for hybrid 2 and 6 locations forhybrid 3) and as the average change in Bu/Ac compared to the base seedtreatment (ST) control treated with surfactant alone in Table 51.

TABLE 51 Foliar treatment using Bt.4Q7Flg22 and a Syn01Flg22 syntheticmutant - increase yield in corn Average Change Application Use Averagein Yield Bu/Ac Foliar Treatment Rate Fl. oz/Ac Yield compared to(Concentration) (mL/hectare) Bu/Ac Surfactant Control Bt.4Q7Flg22 4.0Fl. oz/Ac 201.43 +1.14 (16.7 μM) (292.3 mL/hectare) (Composition 1)Bt.4Q7Flg22 4.0 Fl. oz/Ac 205.43 +2.55 (16.7 μM) + (292.3 mL/hectare)Cellobiose 8.0 Fl. oz/Ac (320 mM) (584.6 mL/hectare) (Composition 4)Syn01Flg22 4.0 Fl. oz/Ac 203.79 +0.90 (16.7 μM) (292.3 mL/hectare)(Composition 6) Syn01Flg22 0.4 Fl. oz/Ac 204.36 +1.48 (16.7 μM)  (29.2mL/hectare) (Composition 7) (16.7 μM) + 0.4 Fl. oz/Ac 204.47 +1.59Cellobiose  (29.2 mL/hectare) (320 mM) 8.0 Fl. oz/Ac (Composition 8)(584.6 mL/hectare)

Corn plants at approximately the V5 stage of development received foliarapplications using a foliar composition comprising a Bt.4Q7Flg22 and asynthetic version Syn01Flg22 of Bt.4Q7Flg22 polypeptides. TheBt.4Q7Flg22 and a synthetic version of Bt.4Q7Flg22 polypeptides werealso combined with cellobiose (320 mM), a reducing sugar, consists oftwo β-glucose molecules linked by a β-(1→4) bond and provided as anelicitor treatment to enhance the effect of the Flg22 polypeptide. Boththe Bt.4Q7Flg22 and the Syn01Flg22 provided in combination withcellobiose to corn plants resulted in an enhanced yield boost over theBt.4Q7Flg22 and the Syn01Flg22 foliar applied polypeptides. A positiveincrease in yield of +2.55 Bu/Ac or 160 kg/Ha resulted in the cornplants that received the Bt.4Q7Flg22 foliar treatment with cellobiose ascompared to the +1.14 BuAc or 71.6 kg/Ha increase in yield for the Bt.4Q7Flg22 foliar treatment provided alone. There was also a positiveincrease in yield of +1.59 Bu/Ac or 99.8 kg/Ha resulted in the cornplants that received the Syn01Flg22 (0.2 Fl. oz/Ac or 14.6 mL/Ha) foliartreatment provided in combination with cellobiose as compared to the+1.48 BuAc or 92.9 kg/Ha increase in yield for the Syn01Flg22 foliartreatment provided at the same application use rate. Whereas, theBt.4Q7Flg22 and the Syn01Flg22 provided as foliar treatments to cornplants at the V5 stage of development using a 4.0 Fl. oz use rate or292.3 mL/Ha provided a slightly lower increase in yield +1.14 Bu/Ac(71.6 kg/Ha) and +0.90 Bu/Ac (56.5 kg/Ha) as compared to thecombinations of the two Flg22 polypeptides with cellobiose.

Example 35: Combination of a Synthetic-Derived Flg22 (Syn01Flg22) and aFungicide

In a further study, large acre yield trials were conducted using afoliar application comprising a compositions of the Bt.4Q7Flg22polypeptide and a synthetic derived polypeptide from Bt.4Q7Flg22(Syn01Flg22) provided with a broad-spectrum fungicide, STRATEGO YLD(10.8% prothioconazole and 32.3% thiofloxystrobin). STRATEGO YLD is acommercially available fungicide suitable for use as an early seasonfoliar application for corn was applied as a foliar spray following therecommendations on the specimen label at a use rate of 4.0 fluid ouncesper acre (Fl. oz/Ac) (292.3 mL/hectare). Corn plants at approximatelythe V5 stage of development received foliar applications using a foliarcomposition comprising the Bt.4Q7Flg22 polypeptide and Syn0Flg22, thesynthetic version of Syn01Flg22 polypeptide combined with the STRATEGOYLD fungicide. Foliar treatments were applied to 2 corn hybrids (DEKALBhybrids: hybrid 1: DKC 52-61; hybrid 2: DKC 58-89) planted in 2locations Iowa. Corn yield (Bu/Ac) was collected and reported as theaverage yield (Bu/Ac) across the 2 locations for both hybrids and as theaverage change in Bu/Ac compared to the corn plants grown from seed thatreceived the base seed treatment (ST) and only the foliar applicationwith the STRATEGOYLD fungicide (Table 52).

TABLE 52 Corn yield foliar applications of a synthetic mutant ofBt.4Q7Flg22 combined with a fungicide Average Change Application UseAverage in Yield Bu/Ac Foliar Treatment Rate Fl. oz/Ac Yield Compared to(Concentration) (mL/hectare) Bu/Ac Fungicide Control STRATEGO YLD 4.0Fl. oz/Ac 223.13 — Fungicide (292.3 mL/hectare) STRATEGO YLD 4.0 Fl.oz/Ac 228.62 +5.49 Fungicide + (292.3 mL/hectare) Bt.4Q7Flg22 4.0 Fl.oz/Ac (SEQ ID NO: 226) (292.3 mL/hectare) (16.7 μM) (Composition 3)STRATEGO YLD 4.0 Fl. oz/Ac 228.96 +5.83 Fungicide + (292.3 mL/hectare)Syn01Flg22 4.0 Fl. oz/Ac (SEQ ID NO: 571) (292.3 mL/hectare) (16.7 μM)(Composition 6) The base seed treatment (ST) consisted of EVERGOLfungicide + PONCHO/VOTIVO 500. The STRATEGO YLD fungicide was applied atthe concentration and application use rate as recommended on thespecimen label.

Foliar application to V5 corn plants with the Bt.4Q7Flg22 and theSyn01Flg22 polypeptides that were provided in combination with afungicide, STRATEGO YLD at the concentrations and application use ratesas specified in Table 5 above resulted in a more than a +5 Bu/Ac. TheSyn01Flg22 polypeptide foliar treatment resulted in slightly higher cornyields of +5.84 Bu/Ac (366.6 kg/Ha) than the corn plants that receivedthe Bt.4Q7Flg22 polypeptide treatment which resulted in average yieldsof +5.50 Bu/Ac (345.2 kg/Ha) as compared to the plants that received thefoliar treatment with only the STRATEGO YLD fungicide.

Example 36: Seed Treatment with Flg22 Polypeptides to Increase Yield inCorn

In other studies, large acre yield trials were conducted using a baseseed treatment consisting of ®EVERGOL fungicide (7.18% propiconazole,3.59% penflufen and combined with 5.74% metalaxyl) and PONCHO/VOTiVO 500(a mixture of 40.3% clothianidin insecticide and 51.6% Bacillus firmusI-1582, a microbial agent) provided in combination with various Flg22polypeptides. Seed treatments were applied to 3 corn hybrids (BECK's4919V2, 5140HR and 5828YX) planted in 8 locations throughout the USMidwest (IN, IL, & IA). Seed treatment compositions of the Flg22polypeptides were applied as described in Table 53 as Fl. oz per unit ofcorn or soy seeds in a total slurry volume containing the base seedtreatment Bt.4Q7Flg22 from Bacillus thuringinesis (Composition 10) andPa. Flg22 from Paenibacillus alvei (Composition 11). Final concentrationof polypeptide in the slurry for Compositions 10 and 11 was 1 uM.

TABLE 53 Compositions of Flg22 seed treatments for testing on corn andsoybean Application Use Rate Fluid ounce/unit corn or soy (Fl. oz/unit)Composition Seed Treatment Formulation Milliliters/unit (mL/unit)Composition 10 Bt.4Q7Flg22 (SEQ ID NO: 226) 40.0 μM 0.14 Fl. oz/unit or11.6 mM Sodium Phosphate Dibasic 4.14 mL/unit combined with 4.2 mMCitric Acid Monohydrate pH 5.6 Composition 11 Pa.Flg22 (SEQ ID NO: 293)40.0 μM 0.14 Fl. oz/unit or 11.6 mM Sodium Phosphate Dibasic 4.14mL/unit combined with 4.2 mM Citric Acid Monohydrate pH 5.6

Corn yield (Bu/Ac) was collected and reported as the yield (Bu/Ac)across the 8 locations averaged for all 3 hybrids. The average change inBu/Ac was as compared to the corn plants grown from seed that receivedthe only the base seed treatment (ST) and is reported in Table 54.

TABLE 54 Corn seed treatment with Flg22 Polypeptide increases yieldAverage Change Average in Yield (Bu/Ac) Application Yield compared toFoliar Treatment Use Rate (Bu/Ac) ST control Bt.4Q7Flg22 0.14 Fl.oz/unit or 179.72 +4.73 Bacillus thuringinesis 4.14 mL/unit (SEQ ID NO:226) (Composition 10) Pa.Flg22 0.14 Fl. oz/unit or 182.24 +3.57Paenibacillus alvei 4.14 mL/unit (SEQ ID NO: 293) (Composition 11)

Treatment of corn seed with Bt.4Q7Flg22 (SEQ ID NO: 226) and Pa.Flg22(SEQ ID NO: 293) polypeptides increased yield as represented as anaverage over the 3 corn hybrids and the 8 US Midwest locations. TheBt.4Q7Flg22 polypeptide provided as a seed treatment resulted in an evengreater yield advantage or a +4.73 Bu/Ac (296.9 kg/Ha) compared to thecontrol plants. The Pa.Flg22 applied as a seed treatment also resultedin a yield gain with a +3.57 Bu/Ac (224 kg/Ha) over corn plants grownfrom seed that received only the base seed treatment. Thus, Flg22polypeptides obtained from different species of bacteria (Bacillus andPaenibacillus) both resulted in substantial yield increases when appliedas a seed treatment on corn seed.

Example 37: Application of Flg22 Polypeptides with Cellobiose toIncrease Yield in Corn

Large acre corn trials were planted from corn seed (DEKALB hybrids: DKC52-61, DKC 58-89, and DKC 65-81) containing a seed treatment comprisingEVERGOL fungicide (7.18% propiconazole, 3.59% penflufen and combinedwith 5.74% metalaxyl) combined with PONCHO/VOTiVO 500 (a mixture ofclothianidin insecticide and a microbial agent, Bacillus firmus 1582).Corn plants at approximately the V5 stage of development received foliarapplications using an agricultural composition comprising a Bt.4Q7Flg22and the synthetic Syn01Flg22 polypeptides were provided with and withoutcellobiose (320 mM). The foliar treatments were applied to 2 cornhybrids (DEKALB hybrids: hybrid 1: DKC 58-89; hybrid 2: DKC 65-81)planted in 2 locations in the US Midwest (IL) that experienceddrought-like conditions after foliar application, during the pollinationstage of corn development. Corn plants received the Bt.4Q7Flg22 andSyn01Flg22 foliar treatments using the concentrations and applicationuse rates as described in Table 48 with a non-ionic surfactant (AlligareSurface™ applied at a final concentration of 0.1% v/v of spray tankvolume). Corn yield (Bu/Ac) was collected and reported as the averageyield (Bu/Ac) across the 2 locations for the 2 hybrids and as theaverage change in Bu/Ac compared to yield from corn plants that receivedonly base seed treatment (ST) and a non-ionic surfactant (AlligareSurface™ applied at a final concentration of 0.1% v/v of spray tankvolume) (Table 55).

TABLE 55 Combinations of Flg22 polypeptides with cellobiose - cornAverage Change Application Use Average in Yield (Bu/Ac) Foliar TreatmentRate Fl. oz/Ac Yield compared to (Concentration) (mL/hectare (Ha)(Bu/Ac) Surfactant control Bt.4Q7Flg22 4.0 96.46 +3.70 (SEQ ID NO: 226)(292.3 mL/Ha) (16.7 μM) Bt.4Q7Flg22 4.0 100.98 +8.22 (SEQ ID NO: 226)(292.3 mL/Ha) (16.7 μM) + 8.0 Cellobiose (584.6 mL/Ha) (320 mM)Bt.4Q7Flg22 48.0  119.37 +26.61 (SEQ ID NO: 226) (3507.6 mL/Ha)  (16.7μM) Syn01Flg22 4.0 98.48 +5.72 (SEQ ID NO: 571) (292.3 mL/Ha) (16.7 μM)Syn01Flg22 0.4 102.36 +9.60 (SEQ ID NO: 571) (29.2 mL/Ha) (16.7 μM)Syn01Flg22 0.4 108.24 +15.48 (SEQ ID NO: 571)  (29.2 mL/Ha) (16.7 μM) +8.0 Cellobiose (584.6 mL/Ha) (320 mM)

Foliar treatment applications of Bt.4Q7Flg22 (SEQ ID NO: 226) and asynthetic version of Syn01Flg22 (SEQ ID NO: 571) resulted in substantialyield gains in corn plants when combined in a foliar treatmentapplication with cellobiose, a disaccharide that is used as a secondarystabilization agent for the Flg polypeptide and vehicle for delivery tothe plant membrane surface. The Bt.4Q7Flg22 polypeptide (16.7 μM)provided with cellobiose (320 mM) as a combination foliar spray appliedusing 4.0 Fl. oz/Ac application use rate (Flg22) resulted in a more thandoubled yield gain, a +8.22 Bu/Ac increase or approximately 516 kg/haover the control plants in comparison to 4.0 Fl. oz/Ac Bt.4Q7Flg22polypeptide alone. The Bt.4Q7Flg22 polypeptide applied withoutcellobiose resulted in a +3.70 Bu/Ac or 232 kg/Ha increase over thecontrol plants grown from the surfactant control. Similar increasedyield resulted in corn plants treated with the Syn01Flg22 and thecombination of Syn01Flg22 (16.7 μM) provided in combination withcellobiose (320 nM) using a 0.2 Fl. oz/Ac application use rate, arespective increase of +9.60 (602.6 kg/Ha) and +15.48 (971.6 kg/Ha)compared to the yield obtained from the surfactant control plants.Additionally, the Bt.4Q7Flg22 (16.7 μM) polypeptide was provided as afoliar spray application using three different application use rates of0.2, 2.0 and 24.0 Fl. oz/Ac (14.6 mL/Ha, 146.2 mL/Ha and 1753.8 mL/Ha)to corn plants at the V5-V7 stage of development. The Bt.4Q7Flg22polypeptide delivered using the highest use rate resulted in asubstantially higher yield advantage, an almost +27 Bu/Ac (1694.6 kg/Ha)yield increase over the yield obtained from the control plants. Overall,Bt.4Q7Flg22 and a synthetic version of Syn01Flg22 provided protectionfrom drought-like growth conditions during a critical stage of plantdevelopment (i.e. pollination), resulting in increased yield for allcombinations of Bt.4Q7Flg22, Syn01Flg22 and cellobiose used as foliarapplications.

In another study, seed treatments using Flg22 polypeptides andcombinations of Flg22 polypeptides with cellobiose resulted in overallyield increases in field trials reported as an average for fourreplicated trials (Table 56). Seed treatments were applied to cornhybrid (BECK's 5828YX) planted in 1 locations in the US Midwest(Columbia). Seed treatment compositions of Flg22 were applied asdescribed in Table 56 as 0.14 Fl. oz per unit of corn seeds in a totalslurry volume containing the base seed treatment. Final concentration ofthe Flg22 polypeptides in the slurry for were standardized to 1 uM perseed. The same final concentration of cellobiose that was applied incombination treatments with the Flg22 polypeptides was at 1.0 mM perseed. The average yield in Bu/Ac and the average increase in Bu/Ac ascompared to the untreated control (column 1) and to the Bt.4Q7Flg22 (SEQID NO:226) (column 2) is reported for corn grown from seed that receivedthe Flg22 polypeptide combination treatments as described below in Table56.

TABLE 56 Seed treatment combinations of Flg22 polypeptides and variantsof Flg22 polypeptides with cellobiose - corn Average Yield Bu/Ac AverageChange Average Change (Average Change in in Bu/Ac in Bu/Ac comparedFoliar Treatment Bu/Ac compared to compared to to Bt.4Q7Flg22;(Concentration) Untreated Control) Untreated Control SEQ ID NO: 226 BaseSeed 28.00 — −1.60 Treatment Control Bt.4Q7Flg22 29.60 +1.60 — at 1.0 μM(SEQ ID NO: 226) Syn01Flg22 39.58 +11.58 +9.98 at 1.0 μM (SEQ ID NO:571) Syn03Flg22 35.17 +7.17 +5.57 at 1.0 μM (SEQ ID NO: 300) Pa.Flg2239.04 +11.04 +9.44 Paenibacillus alvei at 1.0 μM (SEQ ID NO: 293)La.Flg22 37.13 +9.13 +7.53 Lysinibacillus at 1.0 μM (SEQ ID NO: 574)Flg22-B2 36.79 +8.79 +7.19 Bacillus at 1.0 μM (SEQ ID NO: 295) Flg22Combination 45.39 +17.39 +15.79 Syn01Flg22 (SEQ ID NO: 571) + Flg22-B2(SEQ ID NO: 295) + AtFlg22-B4 (SEQ ID NO: 300) at 0.33 μM eachCellobiose 1 mM 39.66 +11.67 +10.07 Bt.4Q7Flg22 36.22 +8.22 +6.62 (SEQID NO: 226) at 1.0 μM + Cellobiose 1 mM Bt.4Q7Flg22 47.06 +19.07 +17.47(SEQ ID NO: 226) at 0.25 μM Bt.4Q7Flg22 40.62 +12.63 +11.02 (SEQ ID NO:226) at 0.25 μM + Cellobiose 1 mM Syn01Flg22 32.65 +4.66 +3.06 (SEQ IDNO: 571) at 0.25 μM Bt.4Q7Flg22 31.03 +3.03 +1.43 (SEQ ID NO: 571) at0.25 μM + Cellobiose 1 mM

Example 38: Application of Flg22 with Cellobiose Additive to V4-V6Soybean Increased Yield—Large Acre Yield Trials

Large acre soybean trials were planted from uncoated soybean seed.Soybean seed was planted 1.5 to 2 inches deep (approximately 5 cm) toensure normal root development. Soybean was planted in 12.5′ (3.8 meter)plots with an average of 150,500 plants per acre, row widths of 30 inchrows (0.8 meter) and seed spacing of 7 to 8 seeds per foot (30 cm).

Agricultural compositions comprising agriculturally effective amounts ofcompositions of Bt.4Q7Flg22 (SEQ ID NO: 226), Syn01Flg22 (SEQ ID NO:571) and a Flg22 from Aneurinbacillus thermoaerophilus, At.Flg22-B4 (SEQID NO: 300) were applied to soybean. The Flg22 polypeptide treatmentswere applied as a foliar spray at application use rates (Fl. oz/Ac ormL/Ha) as specified in Table 57 to soybean grown at five US Midwestlocations (participating sites: IA and IL). The soybean plants receivedfoliar treatments containing Bt.4Q7Flg22 (SEQ ID NO: 226); Syn01Flg22(SEQ ID NO: 571) and At.Flg22-B4 (SEQ ID NO: 300) at approximately theV4-V6 stage of development with a non-ionic surfactant to facilitatespreading and uptake of treatments (Alligare Surface™ applied at a finalconcentration of 0.1% v/v of spray tank volume). Soybean yield wascollected for the 3 soybean varieties (Asgrow: AG2733, AG3536 andAG4034) for plants receiving the Flg22 compositions. Soybean yield wasalso reported as the change in yield Bu/Ac compared to the controlsoybean plants that received a non-ionic surfactant (Alligare Surface™applied at a final concentration of 0.1% (v/v) only treatment (Table57).

Foliar application of the Bt.4Q7Flg22 and Syn01Flg22 polypeptides werealso combined with cellobiose as an additive and examined for the effectof Flg22 polypeptides combined with the cellobiose additive on yieldincrease. Cellobiose is a glucose disaccharide and a building block forcellulose polymer. Chemically, it is glucose-beta-1-4-glucose, areducing sugar that consists of two β-glucose molecules linked by a β(1-4) bond. Cellobiose is obtained by the breakdown of cellulose orlichenin and yields glucose upon hydrolysis. The cellobiose additivecombined with Bt.4Q7Flg22 resulted in an increase in reactive oxygenspecies (ROS) activity in soybean. Soybean yield was collected for the 3soybean varieties (Asgrow: AG2733, AG3536 and AG4034) for plantsreceiving the Flg22 compositions with and without the cellobioseadditive and reported as the average yield (Bu/Ac) for all 3 varietiesacross locations. Soybean yield was also reported as the change in yieldBu/Ac compared to the control soybean plants that received a non-ionicsurfactant (Alligare Surface™ applied at a final concentration of 0.1%v/v only treatment) (Table 57).

TABLE 57 Soybean yield with foliar treatments using varying Flg22polypeptides Change in Application Use Bu/Ac Over Treatment Rate Fl.oz/Ac Average Bu/Ac Surfactant Concentration (mL/hectare (Ha) (5locations) Control Non-ionic Surfactant 0.1% v/v spray 61.36 — aloneBt.4Q7Flg22 4.0 Fl. oz/Ac 62.85 +1.49 (SEQ ID NO: 226) (292.3 mL/Ha)16.7 μM Composition 1 Bt.4Q7Flg22 4.0 Fl. oz/Ac 64.72 +1.56 (SEQ ID NO:226) (292.3 mL/Ha) 16.7 μM Composition 2 Bt.4Q7Flg22 4.0 Fl. oz/Ac 63.87+2.51 (SEQ ID NO: 226) (292.3 mL/Ha) 16.7 μM Composition 3 Bt.4Q7Flg224.0 Fl. oz/Ac 63.15 +1.79 (SEQ ID NO: 226): (292.3 mL/Ha) 16.7 μM + 8.0Fl. oz/Ac Cellobiose: 320 mM (584.6 mL/Ha) Composition 4 Bt.4Q7Flg2248.0 Fl. oz/Ac  62.64 +1.28 (SEQ ID NO: 226) (3507.6 mL/Ha)  16.7 μMComposition 5 Syn01Flg22 4.0 Fl. oz/Ac 63.12 +1.76 (SEQ ID NO: 571)(292.3 mL/Ha) 16.7 μM Composition 6 Syn01Flg22 0.4 Fl. oz/Ac 62.88 +1.52(SEQ ID NO: 571) (29.2 mL/Ha) 16.7 μM Composition 7 Syn01Flg22 0.4 Fl.oz/Ac 63.92 +2.56 (SEQ ID NO: 571) (29.2 mL/Ha) 16.7 μM + 8.0 Cellobiose(320 mM) (584.6 mL/Ha)  Composition 8 AtFlg22-B4 4.0 Fl. oz/Ac 63.66+2.30 (SEQ ID NO: 300) (292.3 mL/Ha)  16.7 μM Composition 9

Foliar treatment of the various Flg22 polypeptides, Bt.4Q7Flg22;Syn01Flg22 and At.Flg22-B4 (Compositions 1-9) all resulted in yieldbenefits when applied on soybean at the V4-V6 stage of developmentcompared to the control soybean plants that were treated with a foliarapplication of surfactant alone. Foliar treatment with Bt.4Q7Flg22(Composition 3) applied at 4.0 Fl. oz/Ac resulted in a +2.51 Bu/Ac(168.8 kg/Ha) increase over control plants (surfactant only). TheSyn01Flg22 (Composition 6) polypeptide applied as a foliar treatmentusing 4.0 Fl. oz/Ac to soybean plants resulted in a yield gain of +1.76Bu/Ac (118.4 kg/Ha) compared to the surfactant only control plants.Syn01Flg22 (Composition 7) and Syn01Flg22 with the cellobiose (320 nM)(Composition 8) applied to soybean plants using a lower application userate of 0.2 Fl. oz/Ac resulted in an increase of +1 Bu/Ac with theaddition of the cellobiose additive or an overall+2.56 Bu/Ac (172.2kg/Ha) increase in yield over the control plants. The At.Flg22-B4(Composition 9) polypeptide applied to soybean (V4-V6) also resulted ina yield benefit of +2.3 Bu/Ac (154.7 kg/Ha) compared to the controlplants or over 3.5 Bu/Ac (235.4 kg/Ha) as compared to plants thatreceived treatment with the non-ionic surfactant only.

In still another study, seed treatments using Flg22 polypeptides andcombinations of Flg22 polypeptides with cellobiose were used as seedtreatments on soybean and resulted in overall yield increases in fieldtrials reported as an average for four replicated trials (Table 58).Seed treatments were applied to 1 soybean hybrid (variety) planted in 1locations in the US Midwest (Columbia, Mo.). Seed treatment compositionsof Flg22 were applied as described in Table 58 as 0.14 Fl. oz per unitof soybean seeds in a total slurry and provided to soybean seed that hada base seed treatment consisting of Poncho VOTiVO 600 FS and EvergolEnergy. The application use rates per each seed treatment were heldconstant at 0.14 Fl. oz/Ac or 4.14 mL/unit. Final concentration of theFlg22 polypeptides in the slurry for were standardized to 1 uM per seed.The same final concentration of cellobiose that was applied incombination treatments with the Flg22 polypeptides was at 1.0 mM perseed. Four replicate plots per each seed treatment were randomized overthe location. The average yield in Bu/Ac and the average change in Bu/Acas compared to the control plants that received only the base seedtreatment are reported in Table 58. The most substantial yield increaseswere seen with Bt.4Q7Flg22 (SEQ ID NO: 226) and Syn01Flg22 (SEQ ID NO:571) when applied as a seed treatment on soybean delivered at a finalconcentration of 1.0 μM of the Flg22 polypeptides and resulting inrespective average yield increases of +5.11 (343.7 kg/Ha) and +9.92(667.1 kg/Ha) over yield from soybean that received the base seedtreatment.

TABLE 58 Seed treatment combinations of Flg22 polypeptides and variantsof Flg22 polypeptides with cellobiose - soybean Average Yield Bu/Ac(Average Change in Average Change in Foliar Treatment Bu/Ac compared toBu/Ac compared to (Concentration) Untreated Control) Untreated ControlBase Seed Treatment 41.11 — Control Bt.4Q7Flg22 at 1.0 μM 46.22 +5.11(SEQ ID NO: 226) Syn01Flg22 at 1.0 μM 51.09 +9.92 (SEQ ID NO: 571)Syn03Flg22 at 1.0 μM 43.61 +2.50 (SEQ ID NO: 573) Pa.Flg22 41.39 +0.28Paenibacillus alvei at 1.0 μM (SEQ ID NO: 293) An,.Flg22 46.01 +4.90Aneurillusbacillus at 1.0 μM (SEQ ID NO: 300) Flg22 Bacillus species44.43 +3.32 (Combination of Flg22 sequences) Syn01Flg22 (SEQ ID NO: 571,Flg22- B2 (SEQ ID NO: 295) & Flg22-B4 (SEQ ID NO: 300) at 0.33 μM eachCellobiose 43.98 +2.87 1 mM Bt.4Q7Flg22 43.40 +2.29 (SEQ ID NO: 226) at1.0 μM + Cellobiose 1 mM Bt.4Q7Flg22 43.80 +2.69 (SEQ ID NO: 226) at0.25 μM Bt.4Q7Flg22 43.52 +2.41 (SEQ ID NO: 226) at 0.25 μM + Cellobiose1 mM

Example 39: Application of RHPP to V5 Corn Increased Yield

Large acre corn trials were planted from corn seed (DEKALB hybrids: DKC52-61, DKC 58-89, and DKC 65-81) containing a seed treatment comprisingEVERGOL fungicide (7.18% propiconazole, 3.59% penflufen and combinedwith 5.74% metalaxyl) combined with PONCHO/VOTiVO 500 (a mixture ofclothianidin insecticide and a microbial agent, Bacillus firmus 1582).Corn plants at approximately the V5 stage of development received afoliar application using an agricultural composition comprising anGm.RHPP polypeptide (SEQ ID NO: 600). The formulated Gm.RHPP polypeptide(Table 59) was applied to the corn hybrids using an application use rateof 8.0 Fl. oz/Ac (584.6 mL/Ha) with 0.1% v/v (of spray tank) non-ionicsurfactant (Alligare Surface™). In total, the trial was conducted at 6locations in the US Midwest (IL, IN, IA), with 1-2 hybrids per locationand 3 replicated plots per hybrid. Corn yield (Bu/Ac) was collected andreported as the average yield (Bu/Ac). The average change in Bu/Ac wascompared to the yield of plants grown from the surfactant control andreported as the combined average yield (Bu/Ac) for the 6 locations (11replicated plots in total) and as overall change in Bu/Ac as compared tocontrol plants. Results are shown in Table 59.

TABLE 59 Foliar treatment using RHPP - increase yield in corn AverageYield Bu/Ac (Average Change in Bu/Ac Foliar Treatment Applicationcompared to surfactant Concentration Rate only control) Surfactantcontrol — 206.15 (Control) Gm.RHPP (SEQ ID: 600) 8.0 Fl. oz/Ac 209.67100 μM (584.6 mL/Ha) (+3.52; 64% win rate) PROXEL BC preservative: 330.7μM; 50.1 μM (CMIT); 21.71 μM (MIT)

Foliar treatment of plants with the Gm.RHPP polypeptide resulted inincreased yields in corn compared to plants that received surfactantalone. The average yield per the 6 locations combined for corn plantsthat received the Gm.RHPP polypeptide foliar treatment was slightly morethan 209 Bu/Ac as compared to 206 Bu/Ac or for the control plants. Theyield for the Gm.RHPP treated plants was increased by 3.52 Bu/Ac or220.9 kg/Ha as compared to the yield from the corn control plants (Table59).

Example 40: RHPP Polypeptide Increases Pod Number in Soybean

Soybean (variety MorSoy) plants were grown from seed with 2 seedsplanted per pot in a controlled environmental growth room underconditions of approximately 300 μmol⁻² s⁻¹ (light photons) for a 13/11light/day cycle and a 21° C. day/15° C. night temperature range untilthe V4 stage of development. Plants were then placed under a long dayconditions consisting of 16/8 light/day cycle and temperature 21-26° C.to promote early flowering and speed up progression to the reproductive(R) growth stage. When the soybean plants had reached the R1 stage ofdevelopment a foliar application containing the Gm.RHPP (SEQ ID NO: 600)polypeptide at a final concentration of 300 nM and a non-ionicsurfactant of 0.10% (NIS90:10; Precision Laboratories, LLC) was appliedto soybean. Soybean plants were provided with the Gm.RHPP formulationand a non-ionic surfactant only control. Both the Gm.RHPP and thenon-ionic surfactant control treatment were applied to 18 plants pertreatment. Six equidistant sprays were provided approximatey 15 cm aboveper each plant for complete coverage of foliage. After treatmentapplication, the R1 soybean plants were returned to the controlenvironmental growth room. After seventeen days, the plants receivedanother foliar treatment application with the formulation containing theGm.RHPP polypeptide and the non-ionic surfactant as well as thenon-ionic surfactant only treatment. Soybean pods of more than 1 mm inlength were counted on the plants after 31 days from the first foliarspray treatment applications. The average number of pods per plant andthe standard deviation from the overall average are reported (Table 60).A p value (p<0.05 for significance) was calculated from a paired T-testcomparison between pod number from plants that received the Gm.RHPP andthe non-ionic surfactant control treatment applications.

TABLE 60 Number of pods in greenhouse grown soybean at 31 days afterfoliar treatment with RHPP Treatment Pod Count Concentration (STDEV)p-value Non-Ionic Surfactant 1.07 0.0116 (NIS90:10 Control) (+0.44)0.01% (v/v) Gm.RHPP (SEQ ID: 591) + 2.00 Surfactant (+1.22) 300 nM *pvalue < 0.05 is statistically significant

Foliar application of the Gm.RHPP polypeptide at early reproductivestage (R1) of soybean plants resulted in an approximately doubled podcount as compared to plants that received the non-ionic surfactantcontrol treatment.

Example 41: Flg22 and RHPP Polypeptides Increase Yield in Tomato andPepper

Foliar application treatments of Bt.4Q7Flg22 (SEQ ID NO: 226) andGm.RHPP (SEQ ID NO: 600) were applied as an exogenous spray at thepre-bloom stage and used to increase yield in tomatoes and jalapenopeppers.

Small scale plots were designed to simulate commercial growingconditions for tomatoes. Tomato plants, variety Roma were started fromtransplants that were grown in a greenhouse for 45 days prior toplanting into 2 raised field row beds with 2 feet (0.6 meters) betweeneach transplant with an average of 30 plants per row bed. Tomatoes weretransplanted three inches beneath the soil surface once the soiltemperature reached 15.6° C. Tomatoes were grown on raised beds coveredwith black plastic mulch. Plants were grown using drip irrigation andfertilizer (80 lbs. or 36.3 kg) nitrogen; 100 lbs. (45.4 kg) phosphate,and 100 lbs. (45.4 kg) potash or potassium) applied following growerguidelines throughout the growing season to provide for optimum plantgrowth and yields. Small raised bed plots were designed to simulate theplanting densities used by commercial growers that generally plant 2,600to 5,800 plants per acre in single rows with 45.7 to 76.2 cm betweenplants in the row on 1.5- to 2-meter centers. [Orzolek et al.,“Agricultural Alternatives: Tomato Production.” University Park: PennState Extension, 2016].

Foliar treatments using Bt.4Q7Flg22 and Gm.RHPP were applied on thetomato plants directly at early bloom (first flower) stage. TheBt.4Q7Flg22 polypeptide foliar composition was applied using anapplication use rate of 4.0 Fl. oz/Ac (292.3 mL/hectare) and the Gm.RHPPpolypeptide foliar composition was applied using an application use rateof 3.2 Fl. oz/Ac (234 mL/hectare) on tomato plants in 10 gallons ofwater per acre with 0.1% v/v non-ionic surfactant (Alligare™ Surface).The Bt.4Q7Flg22 and Gm.RHPP treated plants were compared to the controlplants that received no foliar treatment application. Plants weretreated in replicates of 6 plants, with three replicates per treatment.Effect of the foliar treatments on the yield obtained from tomatoes wasdetermined and reported as normalized to no spray control treatment. Theaverage fruit weight per tomato plant is reported as the combinedaverage for 2 separate harvests and the average percentage change infruit weight as compared to the no-spray control in Table 61.

TABLE 61 Foliar treatment on Spring-planted tomato Percentage ChangeAverage Fruit in Fruit Weight Foliar Treatment Weight (grams) Comparedto No- Concentration per Plant Spray Control No Spray Control 1369.9 —Bt.4Q7Flg22 (SEQ ID NO: 226) 16.7 μM 1487.8 +8.61% 1.67 mM SodiumPhosphate Buffer, pH 5.7 PROXEL BC preservative: 330.7 μM; 50.1 μM(CMIT); 21.71 μM (MIT) (Composition 3) Gm.RHPP (SEQ ID NO: 591) 100 μM1397.1 +1.99% PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM(MIT)

Foliar treatment application with the Bt.4Q7Flg22 polypeptide providedat a concentration of 16.7 μM and an application use rate of 4.0 Fl.oz/Ac (292.3 mL/hectare) resulted in an overall increase in the averagefruit weight per plant as reported in total grams and an +8.61% changein fruit weight as compared to the no spray control. The Gm.RHPPpolypeptide provided at a concentration of 100 μM and a 3.2 Fl. oz/Ac(234 mL/hectare) application use rate also resulted in yield overallincrease in average fruit weight (grams) per plants and an almost+2%change in fruit weight as compared to the no spray control.

In another study, foliar treatments with the Bt.4Q7Flg22 (SEQ ID NO:226) and Gm.RHPP (SEQ ID NO: 600) polypeptides were applied on jalapenopeppers (Capsicum) plants at early bloom (first flower) stage.Small-scale plots were designed to simulate commercial growingconditions for jalapeno peppers. Peppers were grown for 12-weeks in acontrolled growth room and then transplanted outside in 2 raised bedscovered with black plastic mulch that had good water-holdingcharacteristics and in soil having a pH of 5.8-6.6. Jalapeno pepperplants were spaced 14-16 inches (38 cm) apart with 16-24 inches (50 cm)between plants containing approximately 25 plants per row bed. Plantswere grown using drip irrigation and fertilizer applied following growerguidelines throughout the growing season to provide optimum conditionsfor plant growth. The raised bed plots were designed to simulate theplanting densities used by commercial growers that generally plantapproximately plants per acre (5,000-6,500 plants per acre or12,355-16,062 plants per hectare) in double rows 35.6-45.7 cm apart withthe beds spaced 5.0-6.5 feet (1.52-1.98 meters) apart from their centers(Orzolek et al., “Agricultural Alternatives: Pepper Production.”University Park: Penn State Extension, 2010).

Foliar treatments using the Bt.4Q7Flg22 and Gm.RHPP polypeptides wereapplied on jalapeno pepper using application use rates of 2.0 Fl. oz/Ac(146.2 mL/hectare) and 4.0 Fl. oz/Ac (292.3 mL/hectare) for Bt.Flg22 and3.2 Fl. oz/Ac (234 mL/hectare) for the Gm.RHPP polypeptide in a sprayvolume of 10 gallons of water per acre with 0.1% v/v non-ionicsurfactant (Alligare™ Surface). Plants were treated in replicates of 6plants, with three replicates per treatment. Replicates with averageyield per plant 50% above or 50% below the median yield for the trialwere excluded as outliers. The Bt.4Q7Flg22 and Gm.RHPP polypeptidefoliar treatments applied on jalapeno pepper plants were compared toplants sprayed with 10 gallons of water per acre with 0.1% v/v non-ionicsurfactant (Alligare™ Surface) alone.

Effects of the Bt.4Q7Flg22 and Gm.RHPP polypeptides used as foliar sprayapplications on pepper yield were determined for two separate harvestsusing a once over harvest approach. The number of peppers and the aboveground biomass per plant were normalized to the yield and to the biomassof the pepper control plants that were treated with surfactant alone(Table 62).

TABLE 62 Foliar treatment on Spring-planted Jalapeno pepper AverageFruit Percentage Change in Weight (grams) Fruit Weight Compared FoliarTreatment and Rate per Plant to Surfactant Control Surfactant control(Alligare^( ™) Surface; 0.1% 123.7 — v/v of spray volume) Bt.4Q7Flg22(SEQ ID NO: 226) 16.7 μM 184.1 +49% 1.67 mM Sodium Phosphate Buffer, pH5.7 PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT)(Composition 3) 2 fl oz/Ac Bt.4Q7Flg22 (SEQ ID NO: 226) 16.7 μM 173.7+40% 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BC preservative:330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 3) 4 fl oz/AcGm.RHPP (SEQ ID NO: 591) 100 μM 156.6 +27% PROXEL BC preservative: 330.7μM; 50.1 μM (CMIT); 21.71 μM (MIT) 3.2 fl oz/Ac.

The Bt.4Q7Flg22 polypeptide applied as a foliar spray application toJalapeno pepper at the pre-bloom stage resulted in substantial increasesin average fruit weight per plant, a +49% increase for 2 Fl. oz/Ac(146.2 mL/hectare) and +40% increase for 4 Fl. oz/Ac (292.3 mL/Ha) ascompared to the surfactant only control plants. The Gm.RHPP polypeptidetreatment also applied as a foliar spray at the pre-bloom stage alsoresulted in an increased average fruit weight in Jalapeno peppers perplant with a +27% increase in the weight of peppers as measured on a perplant basis as compared to the peppers harvested from the surfactantonly control plants.

Example 42: Application to Squash—Increased Yield

Foliar treatments containing the Bt.4Q7Flg22 or the Gm.RHPP polypeptidewas applied exogenously as a foliar treatment to Crookneck squash at thefirst bloom stage. Foliar treatments with the Bt.4Q7Flg22 and theGm.RHPP polypeptide were applied to squash plants using an applicationuse rate of 2.0 Fl. oz/Ac (146.2 mL/hectare) or 3.2 Fl. oz/Ac (234mL/hectare), respectively, in a spray volume of 10 gallons of water peracre with 0.1% v/v non-ionic surfactant (Alligare™ 90). Yieldcomparisons were made between the plants treated with the polypeptidescompared to surfactant only control plants, with three replicates pertreatment. Yield for the foliar treated plants that received theBt.4Q7Flg22 or Gm.RHPP polypeptide treatment are reported in Table 63 asthe average weight (grams) of squash per plant over two harvests perreplicate and represented as a percentage change as compared to controlplants. Replicates with average yield per plant 50% above or 50% belowthe median yield for the trial were excluded as outliers.

Squash plants were cultivated in sandy loam soil as follows. 2.5 cmholes were cut in 0.76 meters wide plastic covered mounds, two rows permound, holes spaced 0.46 meters apart within each row. Rows werestaggered within the mound. Mounds were spaced 1.2 meters apart. Threesquash seeds were planted per hole and thinned to a single plant perhole 14 days after planting. Drip irrigation tubing was laid in thecenter of each mound, and plants were watered as necessary.

TABLE 63 Foliar treatment with a composition of Gm.RHPP polypeptide toincrease yield in squash Percentage Change in Average squash fruit FruitWeight Compared weight (grams) per to Surfactant only Foliar Treatmentand Rate plant control Surfactant control (Alligare ™ 90; 716.7 — 0.1%v/v of spray volume) Bt.4Q7Flg22 (SEQ ID NO: 226) 16.7 μM 748.4 +4.4%1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BC preservative: 330.7μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 3) 2 fl oz/Ac Gm.RHPP(SEQ ID NO: 591) 100 μM 748.4 +4.4% PROXEL BC preservative: 330.7 μM;50.1 μM (CMIT); 21.71 μM (MIT) 3.2 fl oz/Ac.

Foliar treatment with either Bt.4Q7Flg22 or Gm.RHPP polypeptide onsquash plants at the pre-bloom stage both resulted in an increasedweight of harvested squash fruit by an average by 31.7 grams per plantor +4.4% change in fruit weight as compared to the surfactant onlycontrol plants (Table 63).

Example 43. Flg22 Polypeptide Reduces Severity of White Leaf Spot onKale

In a replicated Fall season kale trial in the Midwest (Columbia, Mo.),very wet and warm growing conditions led to the development of whiteleaf spot on the kale leaves, which is typically caused by Cercosporabrassicicola. The infected kale plants had received no previous foliartreatments for fungal disease prevention. To assess the severity ofdisease, a scoring rubric (1-5 scale) was established where 1=a healthyplant with three or fewer white fungal spots, 2=a plant with more fouror more spots and a portion of the foliage is affected by disease,3=majority of the foliage shows symptoms and up to one leaf has fallenoff due to disease, 4=majority of the foliage shows symptoms and 2-3leaves have fallen off due to disease, and 5=majority of the foliageshows symptoms and four or more leaves have fallen off due to disease. Asingle person scored all the plants within the trial area, and thenevenly distributed the plants by disease score between the treatments inTable 64, with 6 replicated blocks of 6 plants per treatment (total=36plants per treatment). To test Bt.4Q7Flg22 (SEQ ID NO: 226) forimprovement of disease symptoms on kale, treatments were applied as afoliar spray at the indicated rates in Table 64 in a carrier volume of10 gallons of water per acre with 0.1% v/v non-ionic surfactant(Alligare™ Surface). Three weeks after foliar treatments, the plantswere scored used the same disease severity rubric. The change in diseasescore was calculated for each plant, and the average change in diseasescore was determined per treatment. Plants were harvested four daysafter assessing disease severity, and yield was measured as plant weight(grams). Outlying values with weights that were either 50% below or 50%above the median weight for the trial were excluded from the dataset.

TABLE 64 Foliar kale treatments for amelioration of white leaf spot.Yield (Average Application Use plant weight in Foliar Treatment Rate Fl.oz/Ac grams) Relative Average Change (Concentration) (mL/hectare) tocontrol (%) in Disease Score Surfactant only control n/a 14.6 0.6 point(100%)  improvement Bt.4Q7Flg22 (SEQ ID NO: 226) 12.0 Fl. oz/Ac 15.1 1.1point (100 μM) 1.67 mM Sodium (876.9 mL/hectare) (103%)  improvmentPhosphate Buffer, pH 5.7 PROXEL BC preservative: 330.7 μM; 50.1 μM(CMIT); 21.71 μM (MIT) Liquid Copper Fungicide Label rate 11.5 1.1 point(54.45 Fl oz/Ac) (79%) improvment Liquid Copper Fungicide + Label Rate12.4 0.8 point Bt.4Q7Flg22 (SEQ ID NO: 226) (54.45 Fl oz/Ac) + (85%)improvment (100 μM) 1.67 mM Sodium 12.0 Fl. oz/Ac Phosphate Buffer, pH5.7 (876.9 mL/hectare) PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT);21.71 μM (MIT)

Foliar treatment of infected kale plants with formulated Bt.4Q7Flg22(SEQ ID NO: 226) led to an improvement in yield and disease symptomsover the control (Table 64). While untreated controls had an averageplant weight of 14.6 g, plants receiving foliar Bt.4Q7Flg22 had anaverage plant weight of 15.1 g and an average improvement in diseasescoring of 0.5 points over control. The application of copper fungicideimproved plant disease scores to the same extent as Bt.4Q7Flg22, yetdecreased yield by 21% (11.5 g) compared to the control. The combinationtreatment of copper fungicide with Bt.4Q7Flg22 increased yield to 12.4 gper plant, but overall Bt.4Q7Flg22 alone gave the greatest yield andplant health benefit in the trial. In conclusion, Flg22 polypeptides canused to slow the progression of fungal infections in vegetables andincrease yield under stressful growing conditions.

Example 44: ROS Screening Assays to Determine Compatibility of Flg22Polypeptide with Seed Treatments

Seed treatments were examined for compatibility with the production ofapoplastic reactive oxygen species (ROS) in corn petiole tissues.Various commercially available seed treatments were examined forcompatibility with the Flg22 polypeptide (Bt.4Q7Flg22; SEQ ID NO: 226)shown to increase yield when applied alone as a seed treatment on corn.ROS activity assays were conducted using corn petiole samples from cornhybrid 5828 YX as described in Example 15 with the exception thatRelative light units (RLUs) were recorded with a SpectraMax Lluminometer (0.5 s integration; 2.0 min intervals) over a time course of40 minutes. Varying concentrations of Bt.4Q7Flg22 (0 and 1000 μM) werecombined with three commercial seed treatments consisting of PPST 2030(a combination of bacteria, Bacillus subtilis 5×108 cfu/mL and Bacilluspumilis 5×108 cfu/mL), ILEVO (48.4% fluopyram) and PONCHO/VOTiVO (amixture of 40.3% clothianidin and a microbial agent, Bacillus firmusI-1582) and tested for the presence of a ROS response in corn petioles.All three seed treatments as described were applied using theapplication use rates per seed as recommended on the individual specimenlabel for each seed treatment. A standard curve was generated usingvarying concentrations of the Bt.4Q7Flg22 polypeptide and resulted in alogarithmic correlation between the RLU and concentration of Flg22 withan R2 of 0.90. The RLU values are the average of 4 separate measurements(4 treatment wells on each plate) and the increase in overall ROS (RLU)(times increase over the background) are shown in parentheses (Table65).

TABLE 65 Seed treatment compatibility with Flg22 polypeptide using ROSassay 1 μM 1 μM Bt.4Q7Flg22 Bt.4Q7Flg22 (SEQ ID NO: 226) (SEQ ID NO:226) (Full-strength ST) (1:10 dilution of ST) Seed (Fold increase (×)(Fold increase (×) Treatment Background over background) overbackground) PPST 2030 15952.2 109313.7  96129.9 (6.9×) (6.0×) ILeVO84716.9 548686.2 382365.1 (6.5×) (4.5×) PONCHO/ 17379.7 120788.9267720.2 VOTiVO (6.9×) (15.4×)

ROS production as measured by RLUs were increased with the addition of 1μM Bt.4Q7Flg22 when combined with each of the seed treatments asdescribed in Table 65. The ROS production (RLU values) with the 1:10dilution of the seed treatments with the addition of 1 μM Bt.4Q7Flg22was also increased as compared to the back ground RLU level for the seedtreatment only or no Bt.4Q7Flg22 polypeptide. The diluted PONCHO/VOTiVOseed treatment combined with 1 μM Bt.4Q7Flg22 was increased more than15× compared to the background or 2.2× compared to the non-dilutedPONCHO/VOTiVO treatment applied per seed following the recommendation onthe specimen label. Therefore, the Flg22 polypeptide is detectable byROS assay when combined with standard seed treatment base at labelrates. When combining such Flg22 polypeptides with a particular seedtreatment, adjustment of either the polypeptide concentration or theseed treatment concentration can be taken into consideration to ensurean optimal ROS response in the plant. These demonstrate the activity ofthe Flg22 polypeptides on plants in the presence of other seed treatmentpackages on the market today.

Example 45: Combinations of Flg22 and FlgII-28 Peptides to Increase ROSActivity in Tomato

In a separate study, the Flg22 and FlgII-28 polypeptides derived fromdistinct regions of flagellin protein were tested separately and incombination for compatibility of response in tomato leaves. While Flg22and FlgII-28 are both microbe-associated molecular patterns (MAMPs) theymay be recognized distinctly by the Flagellin-sensing 2 (FLS2) andFlagellin-sensing 3 (FLS3) receptors, respectively (Hind et al., 2016;Nature Plants 2:16128), and the interactions may differ across plantspecies. Several Flg22 polypeptides (Bt.4Q7Flg22, SEQ ID NO: 226;Bt.4Q7Flg22-Syn01, SEQ ID NO: 571 and Ec.Flg22, SEQ ID NO: 526) werecompared using ROS activity assays in tomato to several FlgII-28polypeptides (Ps.tomatoFlgII-28, SEQ ID NO: 751; A.sp.FlgII-28, SEQ IDNO: 375).

Tomato leaves were excised from 4-week-old plants using a cork borer togenerate 4 mm disks. Each disc was cut in half using the edge of a razorblade, and then each disc half was floated on 150 μL of water in a96-well plate to rest overnight. The next day, the water was removedfrom each well just prior to polypeptide treatment. The Flg polypeptidesas described in Table 66 were added to water to bring them to a finalconcentration of 5 nM (Table 67) and 100 nM (Table 68) in solution withluminol and HRP before adding to each treatment well. To maintainactivity, the polypeptides were stored in small aliquots to avoidmultiple freezing and thawing. All dilutions to obtain workingconcentrations were done in ultrapure water. Polypeptide solutions werestored at −20° C. for short term usage or −80° C. for long term storage.RLU values and relative ROS activity (Tables 67, 68) is reported as theaverage of 4 measurements. ROS activity assays were conducted using themethods as previously reported in Example 15 with the exception thatRelative light units (RLUs) were recorded with a SpectraMax Lluminometer (0.5 s integration; 2.0 min intervals) over a time course of40 minutes.

TABLE 66 Flg22 and FlgII-28 Polypeptides from various sources AminoFlg Polypeptide Acid Description Length Sequence Bt.4Q7Flg22 22DRLSSGKRINSASD Bacillus DAAGLAIA thuringiensis (SEQ ID NO: 226)Syn01Flg22 22 DRLSSGKRINSAKD Synthetic DAAGLAIA (SEQ ID NO: 571)Ps.tomato FlgII-28 28 ESTNILQRMRELAV Pseudomonas QSRNDSNSATDREAsyringae pv.  Tomato DC3000 (SEQ ID NO: 751) Ec.Flg22 22 ERLSSGLRINSAKDEscherichia coli DAAGQAIA (J26) (SEQ ID NO: 526) A.sp.FlgII-28 28EIHEMLQRMRELAV Aneurinibacillus QAANGTYSDKDKKA sp. XH2 (SEQ ID NO: 300)

TABLE 67 Comparison of ROS activity of Flg22 and Flgll-28 polypeptidesin tomato leaf tissue Average RLU value (5 nM Flg polypeptide) (Foldincrease (×) over Polypeptide Treatment Bt.4Q7Flg22 treatment) Negativecontrol (water) 24423 (0.7×) Bt.4Q7Flg22 33118 (—)  Bacillusthuringiensis (SEQ ID NO: 226) Bt.4Q7Flg22-Syn01 11675 (3.5×) Synthetic(SEQ ID NO: 571) Ps.tomatoFlgll-28 1019995 (30.8×)  Pseudomonas syringaepv. Tomato (SEQ ID NO: 751) Ec.Flg22 426307 (12.9×) Escherichia coli(SEQ ID NO: 526) Aneurinibacillus. sp. Flgll-28 32980 (1.0×) (SEQ ID NO:375)

TABLE 68 Flgll-28 polypeptides from gram-negative Pseudomonas syringaepv. Tomato DC3000 and gram-positive Aneurinibacillus sp. XH2 trigger ROSproduction in tomato leaf tissue Average RLU value with 100 nM Flgpolypeptide Polypeptide Treatment (Fold increase (×) over ConcentrationBt.4Q7Flg22 treatment) Negative control (water)    15,824 (0.007×)Bt.4Q7Flg22 2,118,932 (—)  Bacillus thuringiensis (SEQ ID NO: 226) 100nM Ps.tomatoFlgll-28 3,657,810 (1.7×) Pseudomonas syringae pv. TomatoDC3000 (SEQ ID NO: 751) 100 nM Bt.4Q7Flg22 4,222,426 (2.0×) (SEQ ID NO:226; 100 nM) + Ps.tomatoFlgll-28 (SEQ ID NO:751; 100 nM)Aneurinibacillus. sp. Flgll-28 2,844,947 (1.3×) (SEQ ID NO: 375) 100 nM

It was determined from the results in Table 67 and Table 68 that asecond epitope of flagellin, termed FlgII-28 derived from eitherGram-negative Pseudomonas syringae pv.tomato DC3000 or Gram-positiveAneurinibacillus sp. XH2 (SEQ ID NO: 375) are sufficient to trigger animmune response (e.g. ROS production) in tomato (SEQ ID NO: 751) at both5 nM and 100 nM concentrations. At the 5 nM concentration, Ps.tomatoFlgII-28 had the highest activity as compared to the other Flg22 andFlgII-28 polypeptides and resulted in an almost 31 times increase inRLUs as compared to Bt.4Q7Flg22 at the same concentration, whereas 5 nMA.spp.Flg1128 gave an equally low ROS response to 5 nM Bt.4Q7Flg22. TheFlg22 polypeptide (Ec.Flg22; SEQ ID NO: 526) from Gram-negativeEscherichia coli also resulted in increased ROS activity when applied totomato leaves, with RLU values 12.9 X over the Bt.4Q7Flg22 treatmentalone. The Bt.4Q7Flg22 (SEQ ID NO: 226) polypeptide triggered a very lowROS response in tomato leaves at the 5 nM concentration, but provided ahigh response at the 100 nM concentration. Ps.tomato FlgII-28, on theother hand, provided a strong ROS response in comparison to the negativecontrol (water) at both tested concentrations. Thus, tomato leavesdisplay increased sensitivity to Flg polypeptides derived fromgram-negative bacteria Flagellin such as Ps.tomato FlgII-28 andEc.Flg22. In addition, a synthetic variant of Bt.4Q7Flg22 termedSyn01Flg22 (SEQ ID NO: 571) had substantially increased activity (3.5×)as compared to Bt.4Q7Flg22 treatment when tested at the 5 nMconcentration.

As indicated in Table 68, combinations of Gram-positive (Bt.4Q7Flg22;SEQ ID NO: 226) and Gram-negative Ps.tomato FlgII-28 (Pseudomonassyringae pv. tomato DC3000; SEQ ID NO: 751) can be used as a combinedfoliar application to increase ROS production over either treatmentalone, and enhance plant immunity against certain pathogenic organisms.

Example 46: Synthetic Flg22Syn01 and Flg-15Syn01 Polypeptides toIncrease ROS Activity in Corn and Soybean

A truncated version of Syn01Flg22 derived from Bt.4Q&Flg22 lacking sevenN-terminal amino acids was generated, resulting in the 15 amino acidpolypeptide with the sequence nh2-RINSAKDDAAGLAIA-cooh. Thispolypeptide, termed Bt.4Q7Syn01Flg15 (SEQ ID NO: 752) is a naturallyoccurring polypeptide among the Gram-negative proteobacteria but isabsent from Gram-positive protein sequences. The core sequence requiredfor receptor interaction, RINSAKDD, is retained in the shortenedpolypeptide, and thus the 15-amino acid variant was predicted to beactive for triggering ROS production in plants. To test this, Syn01Flg15was compared to Bt.4Q7Flg22 and Syn01Flg22 in ROS assays with both corn(Table 69) and soybean (Table 70). ROS activity assays were conductedusing the methods as previously reported in Example 15 with theexception that Relative light units (RLUs) were recorded with aSpectraMax L luminometer (0.5 s integration; 2.0 min intervals) over atime course of 40 minutes.

TABLE 69 Flg22Syn01 and Flg15Syn01 variants have greater activity thanBt.4Q7Flg22 in a ROS activity assay with corn stalk tissue. FlgPolypeptide Bt.4Q7Flg22 Syn01Flg22 Syn01Flg15 Concentration (SEQ ID (SEQID (SEQ ID (nM) NO: 226) NO: 571) NO: 752) 100 33037 54888 n.d. (1×*) (1.6×) 10  6032 17660 14079 (0.2×) (0.5×) (0.4×) *Relative ROS activitywas normalized to the average RLU values of Bt.4Q7Flg22 (SEQ ID NO:226). n.d. indicates that a value was not tested and therefore arelative value was not determined.

In the ROS activity assay with corn (Table 69), the Flg22□Syn01 (SEQ IDNO: 571) had the greatest ROS response in corn stalk tissue at both the100 nM and 10 nM concentrations as indicated by the relative respectiveactivities of 1.6× (100 nM) and 0.5× (10 nM) as compared to treatmentusing Bt.4Q7Flg22 (SEQ ID NO: 226) that has an attenuated ROS responseof 0.2× at 10 nM. The shortened version of Syn01Flg22 (SEQ ID NO: 571)or Syn01Flg15 (SEQ ID NO: 752) also exhibited a greater ROS response of0.4× at 10 nM, which was twice the relative ROS activity of Bt.4Q7Flg22(SEQ ID NO: 226) at the same concentration.

TABLE 70 Flg22Syn01 and Flg15Syn01 variants have greater activity thanBt.4Q7Flg22 in a ROS assay with soybean leaf tissue Relative ROSRelative ROS Relative ROS Activity Activity Activity Flg PolypeptideBt.4Q7Flg22 Flg22Syn01 Syn01Flg15 Concentration (nM) (SEQ ID NO: 226)(SEQ ID NO: 571) (SEQ ID NO: 752) 100 250,432 315,961 n.d. (1×)*  (1.25×) 10  10,754  62,020 42,983 (0.04×) (0.25×) (0.17×) *Relative ROSactivity was normalized to the average RLU values of Bt.4Q7Flg22 (SEQ IDNO: 226). n.d. indicates that a value was not tested and therefore arelative value was not determined.

Likewise, in the ROS activity assay with soybean (Table 70), thesynthetic derived mutant of Bt.4Q7Flg22 described as Bt.4Q7Flg22□Syn01(SEQ ID NO: 571) also had the greatest ROS response in soy leaf tissueat both the 100 nM and 10 nM concentrations as indicated by the relativerespective activities of 1.25× (100 nM) and 0.25× (10 nM) as compared totreatment using Bt.4Q7Flg22 (SEQ ID NO: 226) that has a highlyattenuated ROS response of 0.04× at 10 nM. The shortened version ofSyn01Flg22 or Syn01Flg15 also exhibited a greater ROS response of 0.17×at 10 nM, which was four times the relative ROS activity of Bt.4Q7Flg22at the same concentration.

Overall, the Syn01Flg22 had higher ROS activity at both concentrationstested in both corn and soy tissues in comparison to Bt.4Q7Flg22 (SEQ IDNO: 226). The shortened 15-amino acid polypeptide Syn01Flg15 was 2-4×more active than Bt.4Q7Flg22 and only slightly less active than the22-amino acid Syn01Flg22 at 10 nM, indicating that key amino acids foreliciting a plant immune response are retained within the sequence.

Example 47: Chemical Modification to Increase ROS Activity for Flg22Polypeptides

Chemical modifications can be made to Flg22 polypeptides to increaseprotein stability against proteolysis and/or promote a longer durationof activity that can result in greater availability to the FLS2receptor. In general, polypeptide modifications can be utilized to 1)stabilize a polypeptide under adverse conditions or in the presence ofproteases, or 2) provide additional function or molecularcharacteristics to the peptide. Modifications for improved stabilityinclude polypeptide cyclization and alternations at the N- andC-termini. Head-to-tail cyclization (i.e. amide bond formation betweenN-terminal amino and C-terminal carboxyl ends) results in a rigidpolypeptide backbone that resists conformational changes, oftenstabilizing peptide-receptor binding and protecting the polypeptidetermini from exoproteases. Alternatively, modification of thepolypeptide termini can stabilize polypeptides through neutralization(C-terminal amidation) and prevention of N-terminal degradation(N-terminal acetylation). Increased polypeptide solubility and stabilitycan also be conferred through the conjugation of a hydrophilic moleculesuch as polyethylene glycol (PEG).

Such modifications used to stabilize Flg22 polypeptides includePEGylation, cyclization and amidation/acetylation, all of which aredescribed in Table 71. Stabilization of polypeptides using PEGylation iscarried out by linking the polypeptide to polyethylene glycol (PEG).Once linked to the polypeptide, each PEG subunit becomes tightlyassociated with 2 to 3 water molecules, which then function inincreasing the solubility of the polypeptide as well as increasing itsoverall structure to make it less susceptible to proteolytic degradationand more accessible to the membrane FLS2 receptor at the plant surface.Cyclization can also be used to increase the stability of the Flgpolypeptide. Stabilization of a polypeptide can also be obtained usingN-terminal acetylation and C-terminus through amidation where thesemodifications generate a closer mimic of the native protein andtherefore may increase the biological activity of the polypeptide.

TABLE 71 Modified Flg22 polypeptides Peptide Description (ReferenceCode) Modification MW Sequence Bt.4Q7Flg22 Native derived 2229.42nh2 DRLSSGKRI (modified sequence NSASDDAAGLAIA SEQ ID from Bacillusconh2 NO: 226) thuringiensis Bt.4Q7Flg22 N-terminal  229.3 Ac DRLSSGKRIN Mod-1 acetylation SASDDAAGLAIA (modified C-terminal nh2SEQ ID amidation NO: 226) Syn05Flg22 Amino acid 2255.46 Ac DRLSSGKRIN(modified substitution SASDDPAGLAIA SEQ ID (A16P) nh2 NO: 578)N-terminal acetylation C-terminal amidation Syn05Flg22- PEGylation2461    peg4 PEG4 before amide (where x = 4) (modified bondDRLSSGKRINSAS SEQ ID conjugated to DDPAGLAIA  NO: 578) Flg22 conh2Syn05Flg22- Cyclization 2196    Cyc(DRLSSGKRI Cyc Head-to-TailNSASDDPAGLAIA) (modified SEQ ID NO: 578)

The specialized, modified polypeptides as described in Table 71including Syn05Flg22-Syn05 (J36), Syn05Flg22-PEG (J37) andSyn05Flg22-Cyc were synthesized by the University of Missouri MolecularInteractions Core (Columbia, Mo. USA), lyophilized to a dry powder, anddetermined to be of the correct MW and desired purity (>70%) by liquidchromatography-mass spectrometry (LC-MS) and high-performance liquidchromatography (HPLC), respectively. Standard synthesis polypeptidesincluding Bt.4Q7Flg22 (SEQ ID NO: 226) and Bt.4Q7Flg22 Mod-1 (SEQ ID NO:226; J41) were obtained from Genscript (Piscataway, N.J. USA). Alllyophilized polypeptides were resuspended in ultrapure water to a 10 mMconcentration and serially diluted in ultrapure water to the desiredconcentration for testing in soybean and corn ROS assays as describedpreviously in Example 15.

For soybean samples, fully expanded trifoliate leaves were removed fromV1 to V3 stage plants (variety Morsoy). Leaf discs (4 mm) were removedusing a cork borer and then floated on 150 μL of water, abaxial sidedown, overnight before performing the ROS assay previously described.

For corn samples, aerial tissue from V1 to V4 stage corn plants (Beck'shybrid 5828 YX) were prepared as previously described. The 1-mm excisedleaf slices were then floated on 150 uL of water overnight.

ROS activity assays were conducted using the methods as previouslyreported in Example 15 with the exception that Relative light units(RLUs) were recorded with a SpectraMax L luminometer (0.5 s integration;2.0 min intervals) over a time course of 40 minutes. Relative lightunits (RLUs) were first plotted over time using a kinetic time coursefor each concentration tested, followed by integration under the curveto calculate total RLU values produced. Average total RLUs (n=4 samplesper treatment) were then graphed versus polypeptide concentration foreach polypeptide for soybean (Tables 72-73) and corn (Table 74).

A best fit logarithmic or linear regression (R>0.80) was fit to the datafor each treatment. Using the best-fit regression, the polypeptideconcentration required to reach a total RLU production of 15,000 totalRLU (corn) or a 50,000 total RLU (soybean) was calculated for eachpolypeptide and % activity was compared within each data set to thecontrol treatment (Tables 72-74).

TABLE 72 Flg22-Bt modified at the N- and C-termini polypeptides triggerreactive oxygen species production in soybean Polypeptide Concentration(nM) for % Activity (compared to Treatment 5 × 10⁴ total RLU unmodifiedBt.4Q7Flg22 (Code) production (SEQ ID NO: 226) Bt.4Q7Flg22 31.4 100.0%(SEQ ID NO: 226) Bt.4Q7Flg22 Mod-1 29.6 106.14% (SEQ ID NO: 226)

The Flg22 polypeptide concentration required to result in an RLU outputof 50,000 RLU for the Bt.4Q7Flg22S Mod-1 (SEQ ID NO: 226) was less thanthe current Bt.4Q7Flg22 (SEQ ID NO: 226) that has been shown to produceyield gains and impart plant protective qualities to soybean plants.This indicates that the modification of Flg22 by N-terminal acetylationand/or C-terminal amidation does not interfere with polypeptide bindingto the FLS2 receptor, and modifications may be used to produce a moreactive and/or stable version of Flg22 as indicated by the +6% increasein activity of Bt.4Q7Flg22S Mod-1 over Bt.4Q7Flg22 (Table 72).

Novel polypeptides were generated at the University of MissouriMolecular Interactions Core (Columbia, Mo.) with a single amino acidsubstitution (A16P) in comparison to the Bt.4Q7Flg22 (SEQ ID NO: 226)unmodified polypeptide, resulting in the Syn05Flg22 (SEQ ID NO: 578)polypeptide which was amenable to further modification by N-terminalPEGylation Syn05Flg22-PEG (SEQ ID NO: 578) and Head-to-Tail cyclizationSyn05Flg22-Cyc (SEQ ID NO: 578). A soy ROS assay was performed to assessthe effect of these two additional modifications, namely N-terminalPEGylation and Head-to-Tail cyclization to a Flg22 polypeptide, withresults shown in Table 73.

TABLE 73 Modified, synthetic Flg22-Bt polypeptides trigger reactiveoxygen species production in soybean Polypeptide Concentration (nM) %Activity (compared to for 5 × 10⁴ total RLU Syn0Flg22; Treatmentproduction SEQ ID NO: 578) Syn05Flg22 89.8 100.0% (SEQ ID NO: 578)Syn05Flg22-PEG 64.8 138.6% (SEQ ID NO: 578) Syn05Flg22-Cyc 146.9 61.1%(SEQ ID NO: 578)

In a soy ROS assay to compare the relative activities of Syn05Flg22 (SEQID NO: 578) to two modified versions of the polypeptide, the PEGylatedpolypeptide Syn05Flg22-PEG (SEQ ID NO: 578) required substantially lessamount of the polypeptide to achieve a total of 50,000 RLU, whichresulted in an increased activity of +38% as compared to thenon-PEGylated version or Syn05Flg22 (SEQ ID NO: 578). PEGylation of theN-terminus of the peptide increases the hydrophilicity of thepolypeptide and may increase affinity for the peptide-binding pocket ofthe FLS2 receptor. The cyclized version of Syn05Flg22-Cyc (SEQ ID NO:578), however, required more polypeptide provided in the ROS activityassay (+57.1 nM more) compared to the non-cyclized version of Syn05Flg22to reach a total RLU production of 50,000 RLU in the soybean ROS assay.This suggests that the cyclization of the Flg22 polypeptide(Syn05Bt.4Q7Flg22-Cyc) may result in a more rigid polypeptide backbonewith altered binding to the FLS2 receptor, such that more cyclizedpeptide is required to reach an equivalent ROS response. However,increased stability of a cyclized polypeptide in the environment maycompensate for the slight loss in activity.

TABLE 74 Modified, synthetic Flg22-Bt polypeptides trigger reactiveoxygen species production in corn Polypeptide Concentration (nM) for %Activity (compared to 5 × 10⁴ total RLU unmodified Syn05Flg22 Treatmentproduction (SEQ ID NO: 578) Syn05Flg22 19.5 100.0% (SEQ ID NO: 578)Syn05Flg22-PEG 15.0 130.3% (SEQ ID NO: 578)

In a corn ROS assay, the PEGylated version of Syn05Flg22-PEG requiredsubstantially less amount (almost 5 nM less) of the polypeptide toachieve a total of 15,000 RLU, which resulted in an increased activityof +30% as compared to the non-PEGylated version or Syn05Flg22 (Table74).

Modification of Flg22 (Bt) or Syn05Flg22 (Bt) polypeptides by N-terminalacetylation, N-terminal PEGylation, C-terminal amidation, and/orhead-to-tail cyclization produces a peptide that retains activity, asmeasured through ROS assays with corn and soy tissues. Thesepolypeptides could be used to deliver a further stabilized Syn05Flg22(SEQ ID NO: 578) derived polypeptide variant for agricultural uses(either by foliar application, seed treatment, in furrow application,application at transplant, or trunk injection). Cyclization ofSyn05Flg22-Cyc may be used to increase the stability of the polypeptideyet compromised the ROS activity, likely by affecting the affinity ofthe synthetic polypeptide to the membrane FLS2 receptor.

Example 48. Adjuvant Compatibility with Flg22 Polypeptides

Product formulations using Flg22 polypeptides can generally includeantimicrobial biostatic preservatives such as Proxel and surfactants.Therefore, the compatibility of these types of adjuvants were testedusing ROS activity assays to determine the effect in solution on Flg22responsiveness when used in combination with such adjuvants. Proxels ingeneral are broad spectrum biocides for the preservation of manyagricultural based products that protect them against spoilage frombacteria, yeast and fungi. Surfactants in general are also commonly usedin agricultural formulations to improve the penetration of manyagrochemical products into the plant for improved performance. In thisstudy, five different Proxels and two different non-ionic surfactantswere tested in formulations combined with Bt.4Q7Flg22 (SEQ ID NO: 226)for effectiveness in producing a ROS response using a ROS activity assayin soybean leaves. The different Proxel formulations (Lonza) aredescribed below in Table 75. Theses Proxel formulations were mixed with40 μM Flg22 polypeptide at a range of recommended label rates by themanufacturer (Lonza), and then diluted into the ROS assay to a finalpolypeptide concentration of 100 nM and Proxel concentrations indicatedin Table 75. The tested non-ionic surfactants were provided in the ROSassay at a range of recommended label rates by the individualdistributer or manufacturer. The average four sample measurements RLUvalues obtained after performing a ROS assay were collected usingsoybean leaf disks as previously described in Example 15 with theexception that Relative light units (RLUs) were recorded with aSpectraMax L luminometer (0.5 s integration; 2.0 min intervals) over atime course of 40 minutes. The average of these 4 RLU values is reportedin Table 76.

TABLE 75 Different PROXEL additives used as adjuvants in formulationswith polypeptides PROXEL Formulations Chemical Description PROXEL BD20 A20% aqueous dispersion of 1,2-benzisothiazoline-3-one PROXELBC Anaqueous dispersion of a blend of 1,2- benzisothiazoline-3-one(BIT),5-chloro- 2-methyl-4-isothhiazoline-3-one (CIMT) and2-methyl-4-isothiazoline-3-one (MIT) PROXELGXL A 20% aqueous dipropyleneglycol solution of 1,2-benzisothiazoline-3-one PROXELBN An aqueousdispersion of 1,2- benzisothiazoline-3-one and2-bromo-2-nitropropen-1,3-diol PROXEL AQ A solution of1,2-benzisothiazoline- 3-one in water

TABLE 76 RLU output values from ROS activity assays in soybean leavesusing Flg22 polypeptide formulated using different Proxel preservativesTreatment Comparison with and without Average RLU values PROXELpreservative (Fold increase over Concentration negative control) Mock(water) Negative Control 4823 Bt.4Q7Flg22  81887 (17.0×) (SEQ ID NO: 226at 100 nM) (No PROXEL Preservative Added) Bt.4Q7Flg22  89188 (18.5×)(SEQ ID NO: 226 at 100 nM) + PROXEL BD20 (0.0005988%) Bt.4Q7Flg22 105527(21.9×) (SEQ ID NO: 226 at 100 nM) + PROXELBD20 (0.00011976%)Bt.4Q7Flg22 136575 (28.3×) (SEQ ID NO: 226 at 100 nM) + PROXELBC(0.0005988%) Bt.4Q7Flg22  92808 (19.2×) (SEQ ID NO: 226 at 100 nM) +PROXELBC (0.00011976%) Bt.4Q7Flg22 128410 (26.6×) (SEQ ID NO: 226 at 100nM) + PROXELGXL (0.0002994%) Bt.4Q7Flg22 101847 (21.1×) (SEQ ID NO: 226at 100 nM) + PROXELGXL (0.0008982%) Bt.4Q7Flg22  91554 (19.0×) (SEQ IDNO: 226 at 100 nM) + PROXELBN (0.0002994%) Bt.4Q7Flg22 105164 (21.8×)(SEQ ID NO: 226 at 100 nM) + PROXELBN (0.00017964%) Bt.4Q7Flg22 116634(24.2×) (SEQ ID NO: 226 at 100 nM) + PROXELAQ (0.0005988%) Bt.4Q7Flg22 98394 (20.4×) (SEQ ID NO: 226 at 100 nM) + PROXELAQ (0.0035928%)

All Proxel preservative treatments as described in Table 76 werecompatible when used in formulations with the Flg22 polypeptide(Bt.4Q7Flg22; SEQ ID NO: 226) as indicated by the high RLU values(19.0-28.3× fold increase over mock treatment) as comparable to theBt.4Q7Flg2 polypeptide control without a Proxel preservative (17.0× foldincrease over mock treatment).

TABLE 77 RLU output values from ROS activity assays in soybean leaftissues using Flg22 polypeptide formulated using different non-ionicsurfactants Treatment Comparison with and without Average RLU valuesSurfactant (Fold increase over Concentration negative control) Mock(water) 51288 Negative Control Bt.4Q7Flg22 350503 (6.8×) (SEQ ID NO:226) 52.2 nM Equivalent: 4.0 Fl. oz/Ac in 10 gallons water/AcBt.4Q7Flg22 142478 (2.8×) (SEQ ID NO: 226) 52.2 nM + Silwet-L77 (0.025%)Bt.4Q7Flg22 163517 (3.2×) (SEQ ID NO: 226) 52.2 nM + Silwet-L77 (0.10%)Bt.4Q7Flg22 329295 (6.4×) (SEQ ID NO: 226) 52.2 nM + NIS90:10 (0.25%)Bt.4Q7Flg22 295726 (5.8×) (SEQ ID NO: 226) 52.2 nM + NIS90:10 (0.5%)

All surfactant (non-ionic) treatments as described in Table 77 werecompatible when mixed at the indicated concentrations with 52.2 nM Flg22polypeptide (Bt.4Q7Flg22; SEQ ID NO: 226), a polypeptide concentrationequivalent to 4.0 Fl oz/Ac usage rate of Composition 1 (Bt.4Q7Flg22; SEQID NO: 226; 16.7 μM) applied in water at a spray rate of 10 gallons peracre. Fold-increase in ROS production over the mock-treated control werecomparable between the Bt.4Q7Flg2 polypeptide control without asurfactant (6.8× over control) versus Bt.4Q7Flg2 polypeptide withnon-ionic surfactant NIS90:10 applied at 0.25% v/v or 0.5% v/v oftreatment solution (5.8-6.4×), or slightly lower for Bt.4Q7Flg2polypeptide with Silwet-L77 applied at 0.025% v/v or 0.1% v/v oftreatment solution (2.8-3.2×). Silwet-L77 (Helena), a non-ionicorganosilicone surfactant is formulated as a co-polymer that hasenhanced wetting and spreading characteristics when used in aqueoussprays. NIS90:10 (Precision Laboratories) is a low-foaming, non-ionicsurfactant that enhances crop protection and performance by improvingspray solution coverage and penetration of target leaf surfaces. Boththe non-ionic surfactants combined with Bt.4Q7Flg22 permitted ROSproduction in response to the Flg22 polypeptide in target leaf tissues(Table 77), and as such, are compatible with Flg22 polypeptide foliarapplication in the field.

Example 49: Production of Bt.4Q7Flg22 Using Fermentation Methods andActivation by Enterokinase Cleavage for Disease Prevention Trials inPotato, Lentils and Citrus Trees

The Bt.4Q7Flg22 (SEQ ID NO: 226) was provided in a confirmation tostabilize the polypeptide and enhance activity for an alternativeproduction method, namely bacterial fermentation. The Bt.4Q7Flg22polypeptide was combined with an amyQ secretion signal from Bacillusamyloliquefaciens alpha-amylase) fused to glutathione S-transferase(GST) and an enterokinase cleavage tag sequence as described: amyQsecretion signal (Bacillus amyloliquefaciens alpha-amylase) GST(Schistosoma japonicum)_linker_Enterokinase cleavagesite_Bt.4Q7Flg22_stop codon (Table 78).

TABLE 78 Cloning of Bt.4Q7Flg22 with sequences toincrease polypeptide stability and activity DescriptionAmino Acid Sequences amyQ secretion signal MIQKRKRTVSFRLVLMCTLL(Bacillus FVSLPITKTSA amyloliquefaciens) SEQ ID NO: 769 GSTMSPILGYWKIKGLVQPTRLL (Schistosoma japonicum) LEYLEEKYEEHLYERDEGDKSEQ ID NO: 770 WRNKKFELGLEFPNLPYYID GDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVL DIRYGVSRIAYSKDFETLKV DFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD VVLYMDPMCLDAFPKLVCFK KRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK linker GGGGGGS SEQ ID NO: 771Enterokinase cleavage tag DDDDK (Consensus cleavage target for bovineEnterokinase, light chain protease) SEQ ID NO: 772 Bt.4Q7Flg22DRLSSGKRINSASDDAAGLA (SEQ ID NO: 226) IA (Bacillus thuringiensisstrain 4Q7) *DNA used for cloning from the amy E secretion signal, GSTand Bt.4Q7Flg22 (SEQ ID NO: 226) sequences came from internalproprietary libraries; production stain code = H101 (Chloramphenicolresistant)

TABLE 79 Cloning of Syn01Flg22 (SEQ ID NO: 571)with sequences to increase polypeptide stability and activityDescription Amino Acid Sequence amyQ secretion MIQKRKRTVSFRLVLMCTLsignal (Bacillus LFVSLPITKTSA amyloliquefaciens) SEQ ID NO: 769 GSTMSPILGYWKIKGLVQPTRL (Schistosoma LLEYLEEKYEEHLYERDEG japonicum)DKWRNKKFELGLEFPNLPY SEQ ID NO: 770 YIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISML EGAVLDIRYGVSRIAYSKD FETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPD FMLYDALDVVLYMDPMCLD AFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQAT FGGGDHPPK linker GGGGGGS SEQ ID NO: 771 EnterokinaseDDDDK cleavage tag (Consensus cleavage target for bovine Enterokinase,light chain protease) SEQ ID NO: 772 Bt.4Q7Flg22-Syn01DRLSSGKRINSAKDDAAGL (SEQ ID NO: 226) AIA (Bacillus  thuringiensisstrain 4Q7) *DNA used for cloning from the amy E secretion signal, GSTand SynFlg22 (SEQ ID NO: 571) sequences came from internal proprietarylibraries. Production strain code = H114 (Tetracycline resistant)

TABLE 80 Cloning of thionin-like proteinwith sequence for secretion into fermentation growth media DescriptionAmino Acid Sequence amyQ secretion signal MIQKRKRTVSFRLVLMCTLSEQ ID NO: 769 LFVSLPITKTSA (Bacillus amyloliquefaciens) Thionin-likeRTCESQSHRFKGPCSRDSN protein CATVCLTEGFSGGDCRGFR (SEQ ID NO: 650)RRCRCTRPCVFDEK (Synthetic)

The sequences in Tables 78, 79 and 80 were cloned into a standardcloning vector containing an ampicillin selection marker and either achloramphenicol (Cm) or Tetracycline (Tet) selection marker that canreplicate in E. coli and then be transferred to Bacillus subtilis strainK08 for production purposes (Production strain codes:H101=amyQ-GST-EK-BtFlg22, H114=amyQ-GST-EK-BtFlg22-Syn01, andH117=amyQ-Thionin-like). The fermentation production was carried out bystarting an overnight culture in sterile 2XYT media (16 g Bactotryptone, 10 g yeast extract, and 5 g NaCl per liter; pH adjusted to7.0) with 10 μg/mL Cm or Tet, and then diluted into fresh 2XYT mediawith 10 μg/mL Cm or Tet the following day. Productions were performedusing 50 mL (shake flask) or 3 L (glass bioreactor vessel) media volumeswith a constant temperature of 30° C. Larger scale up volumes caninclude 5 L to 1000 L+, including up to 30,000 L volumes). Bacterialgrowth was monitored until the culture reached an optical density of0.6-1.0, after which Isopropyl β-D-1-thiogalactopyranoside (IPTG) wasadded to a final concentration of 0.1-1.0 mM to induce production of theGST-Bt.4Q7Flg22 fusion protein. The induced production continued inculture conditions for an additional 12-24 hours to produce the fusionprotein which is secreted into the growth media. Upon secretion, theamyQ secretion tag is cleaved from the fusion protein. The cultures werethen centrifuged at 5000×g for 20 min and filtered through a 0.22 μmbottle-top vacuum filter to remove the bacterial cells. The sterilefiltrate was then collected and used as a foliar treatment on lentil andpotato plants in Sclerotinia disease prevention trials (Example 50) oras a trunk injection of citrus trees for eradication and prevention ofHLB disease symptoms (Example 51).

After fermentation, two versions of the Bt.4Q7Flg22 (SEQ ID NO: 226)polypeptide product were used in potato and lentil disease preventiontrials, one without enterokinase treatment (H101 filtrate,non-activated) and another with activation using a enterokinase tocleave off the GST tag fused to the Flg22 polypeptide (H101 filtrateEK-activated). To activate Bt.4Q7Flg22, the addition of 32 U (units) ofenterokinase (EK: Enterokinase light chain; New England BioLabs, Inc,Product No. P8070) was added per 1 mL of H101 filtrate with anincubation period for 2-3 hours at 30° C. for the enzymatic release ofBt.4Q7Flg22 from the GST-EK cleavage site resulting in an activated andreleased product comprising the 22 amino acid Bt.4Q7Flg22 polypeptides.For the citrus tree injection trial, H101 filtrate and H114 filtratewere EK-activated with the addition of 0.8 U Enterokinse, light chain(New England Biolabs, Inc, Product No. P8070) per mL of filtrate with anincubation of 3 hours at 30° C. for the enzymatic release of Flg22polypeptide. No activation treatment was required for release of thethionin-like peptide which was produced without a GST tag.

Example 50: Foliar Pre-Treatment with Bt.4Q7Flg22 Polypeptides ProtectLentil and Potato Plants from Sclerotinia Stem Rot (White Mold) Disease

Treatment applications of Bt.4Q7Flg22 (SEQ ID NO: 226) were examined forprotection of lentil and potato plants against disease infection andprogression with Sclerotinia sclerotorium strain MT07 (white mold).Three different versions of the Bt.4Q7Flg22 polypeptides were examinedin the disease assessment studies. A formulated Bt.4Q7Flg22 (100 μM) insodium phosphate buffer, pH 5.7 and two different versions ofBt.4Q7Flg22 produced using fermentation methods as described (Example49) and provided with and without activation of Flg22 with anenterokinase (EK) and referred to as H101 filtrate.

Prior to using the three versions of the polypeptides in diseaseprotection assays with lentil and potato plants, a ROS activity assaywas performed using corn petiole tissues using methods as describedpreviously in Example 15 to ensure that the Bt.4Q7Flg22 H101 filtrate,particularly with the EK was active. The H101 Bt.4Q7Flg22 filtrateswithout and with enterokinase (EK=8 U/mL filtrate) activation werecompared to the synthetic Bt.4Q7Flg22 (SEQ ID NO: 226) which was used togenerate a series of concentration comparisons to predict the Flg22concentrations in the H101 filtrates generated using fermentationprocedures.

TABLE 81 ROS activity assay using Flg22 produced by fermentation withand without enterokinase activation in corn Change (×) in Average RLU ofRLU of Flg22 Flg22 Polypeptide Flg22 Polypeptide Polypeptide ComparedConcentration Treatment to Negative Control Bt.4Q7Flg22 6851 0.8× (SEQID NO: 226) (1.0 nM) Bt.4Q7Flg22 9389 1.1× (SEQ ID NO: 226) (5 nM)Bt.4Q7Flg22 12157 1.4× (SEQ ID NO: 226) (25 nM) Bt.4Q7Flg22 25212 3.0×(fermentation H101 filtrate) With (+) Enterokinase 8 U/mL (0.1% v/v)Bt.4Q7Flg22 16891 2.0× (fermentation H101 filtrate) Without (−)Enterokinase (0.1% v/v) *RLU values are reported as an average of 4separate measurements after the background RLU levels were subtracted.

The fermentation produced H101 filtrates of Bt.4Q7Flg22 provided withand without EK activation both resulted in ROS activities (RLU values)that were higher than the control (0 nM Bt.4Q7Flg22). The H101Bt.4Q7Flg22 filtrates (0.1% v/v) with EK treatment provided to corn stemin the ROS assay resulted in a 3.0× increase in RLU values as comparedto the control treatment without any Flg22 polypeptide and the ROSresponse was greater than Bt.4Q7Flg22 (SEQ ID NO: 226) provided at aconcentration of 25 nM; therefore, the estimated Bt.4Q7Flg22 activity inthe undiluted EK-activated filtrate was 25 μM The fermentation producedH101 filtrates of Bt.4Q7Flg22 treatments that were provided without EKstill had ROS activity (2.0× RLU) over the negative control treatmentbut with a lower increase seen in RLU values as compared to the H101filtrates of Bt.4Q7Flg22 with EK. Once it was confirmed that the H101filtrates had activity in ROS assays (Table 81) they were assessed indisease protection studies with potato and lentil.

Various formulations of Bt.4Q7Flg22 polypeptides were provided to lentiland potato plants as a foliar pre-treatment to plants 48 hours prior toinoculation with the Sclerotinia sclerotorium fungus and provided incombination with and without a fungicide (Endura, active ingredient 70%boscalid), which is effective in the treatment and protection of plantsfrom infection with the Sclerotinia fungus (white mold). The Bt.4Q7Flg22formulations were tested using a crop-disease model (Montana StateUniversity, Extension Services Crop Protection) to examine the effectsof each of the foliar pre-treatments on the prevention and protectionagainst disease and the development of symptoms. All treatmentsincluding the water control were applied to the lentil and potato plantsusing an air brush connected to a regulated air compressor set with anoutput pressure to the brush at 50 psi. After the pre-treatment, plantswere inoculated with Sclerotinia sclerotorium using mycelial plug (agarplug covered with mycelium) placed with the mycelia side touching theplant stem and placed in humidity (100%) chambers for a set amount oftime.

Lentils

Lentil (variety Pennel) plants were grown in soilless media consistingof a mixture of 1:1 peat moss to perlite in 4′4′ pots with one plant perpot for 24 days in a controlled growth chamber under growth conditions:300-400 μmol m⁻² s⁻¹ (light photons) for a 13/11 light/day cycle and a21° C. day/15° C. night temperature range. The disease studies includedfive lentil plants per each of six different foliar treatments with 6replicate plants per treatment, a total of 30 plants per foliartreatment as described in Table 35. All of the foliar treatments usedfor pre-treatments were applied with the addition of a non-ionicsurfactant (ALLIGARE SURFACE; Alligare, LLC) to a final concentration of0.1% (v/v) or a concentration of alkylpolyoxethylene, glycolderivatives. Each of the Bt.4Q7Flg22 treatments from the formulated andfermentation-derived productions were provided at an application userate of 0.1% (v/v) or 300 μL of product to 300 mL water and provided toeach plant in an equivalent number of sprays completely covering thefoliage, using 8 mL of each treatment application for all 30 plants pertreatment. The Endura fungicide pre-treatment was applied at anequivalent application use rate of 11 Fl. oz/Ac (803.8 mL/Ha) followingthe application instructions on the specimen label. The treatments wererandomized using a complete random block design. Approximately 48 hoursafter the pre-treatment, plants were inoculated with a Sclerotiniasclerotorium strain isolated locally in Montana using mycelial plug(agar plug covered with mycelium) placed with the mycelia side touchingthe plant stem. The lentil plants were then placed in humidity (100%)chamber for a period of 72 hours. At 11 days after inoculation diseasesymptoms were assessed and scored and average fresh weight (total weightof each replicate—grams) were collected (Table 82). Plants were allowedto dry for approximately 3 weeks, and then dry weight was collected(total weight per replicate—grams) (Table 82)

Disease scoring (disease scoring scale 0-7) and fresh weight and dryweight (grams) were collected for each replicate of five plants and thenaveraged for the total number of plants (n=30). The disease scoring wasranked on a scale of 0-7, with a score of 0 equivalent to no disease anda score of 10 ranked as all plants did not survive (Table 82).

TABLE 82 Disease assessment in lentils 10-days post infection withSclerotinia sclerotiorum Average Total Average Total Disease FreshWeight Dry Weight Scoring per Plant per Plant Scale 0-7 (grams) (grams)Treatment (STDEV) (STDEV) (STDEV) Water control 2.83 4.52 0.95 (±1.84)(±1.18) (±0.21) Endura Fungicide 0.50 2.89 0.68 (±0.84) (±0.30) (±0.05)Formulated 1.33 5.44 0.99 Bt.4Q7Flg22 (±0.82) (±0.48) (±0.07) EnduraFungicide + 1.0  4.95 0.96 Formulated (±0.89) (±0.59) (±0.05)Bt.4Q7Flg22 H101 filtrate non- 2.17 4.51 0.85 activated (±1.47) (±0.93)(±0.06) H101 filtrate EK 2.17 6.11 1.09 activated (±1.33) (±0.69)(±0.11) *p value of ≤0.1 means there is a statistically significantdifference between treatments and the water control.

Foliar application of formulated Bt.4Q7Flg22 was compared to the Endurafungicide, the Endura fungicide combined with formulated Bt.4Q7Flg22 andthe two Bt.4Q7Flg22 treatments provided with the Flg22 polypeptidesproduced from the fermentation reactions with and without EK activationas previously described in Table 82. All of the foliar treatments in thecrop-disease model were compared to the each other and to water controltreated plants and assessed 11 days post inoculation for the appearanceof disease symptoms. Each plant was assigned a disease score from 0-7.The total fresh and dry weights (grams) were also determined per plant.

The Endura fungicide, a commercially available treatment for Sclerotiniasclerotium resulted in the least disease symptom development on lentilcompared across all of the foliar treatments with a disease score of0.50 whereas, the water treatment (control) resulted in a diseaseranking score of 2.83. Foliar application of the formulated Bt.4Q7Flg22treatment to lentil plants resulted in an increased resistance toSclerotinia with a disease score of 1.33 (p value=0.0972) compared toplants that received the water control treatment. Unlike the Endurafungicide treatment which resulted in slowed growth compared to theplants treated with the water control, the formulated Bt.4Q7Flg22treatment resulted in continued vigorous growth during early symptomdevelopment. The lentil plants that received the pre-treatment with theformulated Bt.4Q7Flg22 had an average fresh weight of 5.44 grams perplant compared to plants treated with the Endura fungicide alone (2.89g) or the water control (4.52 g). The combination treatment of theEndura fungicide with the formulated Bt.4Q7Flg22 polypeptide furtherincreased protection of the lentil plants from symptom development witha disease score of 1.0 (p value=0.0524) compared to the plants treatedwith the water control. Plant weight (fresh and dry) for plants thatreceived the pre-treatment with the formulated Bt.4Q7Flg22 polypeptidewas greater than the fresh or dry weights from plants that received thewater control or the Endura fungicide alone. The Bt.4Q7Flg22polypeptides provided from the fermentation derived products (non-EKactivated and EK activated) were equivalent in the disease symptomranking with a disease score of 2.17, which was less than the diseasescore of plants treated with the water only control application.However, the fresh weight per plant treated with the EK-activatedversion of the Bt.4Q7Flg22 polypeptide had a significantly increasedfresh weight of 6.11 grams (p value=0.0266) as compared to the watertreated plants. The EK-activated version of the Bt.4Q7Flg22 polypeptidealso had the overall highest fresh and dry weights compared to all ofthe other treatments in Table 82. Other significant findings of thisstudy were that the formulated Bt.4Q7Flg22 polypeptide pre-treatment oflentil plants protected the lentils from fungicide-induced damage. Theaverage fresh weight of the plants that received the Endura fungicidewas 2.89 g while the formulated Bt.4Q7Flg22 treatment was 5.44 g (pvalue=2.645×10-05). The fermentation produced Bt.4Q7Flg22 containing theenterokinase (EK) enzyme was used to cleave the Bt.4Q7Flg22 polypeptidefrom the GST-EK-Bt.4Q7Flg22 as previously described. This Bt.4Q7Flg22filtrate treatment provided to lentil increased activity of the Flg22polypeptide thus resulting in significantly enhanced plant growth duringthe infection period compared to the water treated control plants (pvalue=1.180×10-05). The non-activated EK or GST-EK-Bt.4Q7Flg22 ornon-cleaved Bt.4Q7Flg22 filtrate did not increase plant growth comparedto the plants that received the water control only treatment (pvalue=0.9852).

Potatoes

Seed potatoes (variety: Russet Burbank) were planted from 2 cm potatosections from which eye buds protrude (1 section per pot) with the cutside down and planted approximately 7-8 cm deep in soilless mediaconsisting of a mixture of 1:1 peat moss to perlite in 10×10 cm pots.Potatoes were grown with one plant per pot for 19 days in a controlledgrowth chamber under standard conditions of receiving approximately300-400 μmol m⁻² s⁻¹ (light photons) for a 13/11 light/day cycle and a21° C. day/15° C. night temperature range. 19 days after planting, thepotato plants were pre-treated with the foliar applications as describedin Table 36. The disease studies included five potato plants per each ofsix different foliar treatments with 6 replicate plants per treatment, atotal of 30 plants per foliar treatment as described in Table 83. All ofthe foliar treatments used for pre-treatments were foliar applied withthe addition of a non-ionic surfactant (ALLIGARE SURFACE; Alligare LLC)to a final concentration of 0.1% (v/v) or a concentration ofalkylpolyoxethylene, glycol derivatives. Each of the Bt.4Q7Flg22treatments from the formulated and fermentation derived productions wereprovided at an application use rate of 0.1% (v/v) or 300 μL of productto 300 mL water and provided to each plant in an equivalent number ofsprays completely covering the foliage using 15 mL of each treatmentapplication for all 30 plants per treatment. The Endura fungicidepre-treatment was applied at an equivalent application use rate of 11Fl. oz/Ac (803.8 mL/Ha) following the application instructions on thespecimen label. The treatments were randomized using a complete randomblock design. Approximately 48 hours after the pre-treatment, plantswere inoculated with Sclerotinia sclerotorium using mycelial plug (agarplug covered with mycelium) placed with the mycelia side touching theplant stem and placed in a humid chamber (100%) for 192 hours. At 16days after inoculation disease symptoms were assessed and scored andaverage stem fresh weight (total stem weight—grams) were collected(Table 83). Plants were allowed to dry for 12 days, and then dry weightswere recorded (total stem weight—grams) (Table 83).

After 48 hours, the potato plants were inoculated with mycelia plugsplaced on the soil near each plant and placed in a humid mistingchamber. The treatments were randomized using a random block design.Disease scoring (scoring scale 0-6). Stem fresh and dry weight (grams)were also collected from each plant and then averaged for the totalnumber of plants (n=30). Stem dry weight was taken after the plants werefully desiccated at approximately 12 days after harvest. Disease scoreswere assessed 16 days after the initial inoculation. The disease scoringwas ranked on a scale of 0-6, with a score of 0 equivalent to no diseaseand a score of 6 ranked as all plants did not survive.

TABLE 83 Disease assessment in potatoes 15-days post infection withSclerotinia sclerotiorum Average Average Fresh Stem Dry Stem DiseaseWeight per Weight per Scoring Plant Plant Scale 0-6 (grams; “g”) (grams;“g”) Treatment (STDEV) (STDEV) (STDEV) Water control 2.83  96.01 12.08(±1.33) (±17.05) (±2.75) Endura Fungicide 0.50 110.98 16.86 (±0.84)(±18.32) (±8.69) Formulated 1.83 113.37 15.01 Bt.4Q7Flg22 (±0.98)(±14.58) (±3.72) (SEQ ID NO: 226) H101 filtrate non- 2.33  98.08 12.96activated (±1.03) (±15.34) (±2.77) H101 filtrate EK 1.83 117.76 17.66activated (±0.75) (±15.83) (±8.70) *p value of ≤0.1 means there is astatistically significant difference between treatments and the watercontrol.

Foliar pre-treatment applications using the formulated Bt.4Q7Flg22 andBt.4Q7Flg22 polypeptides derived from the fermentation products (H101filtrates) were compared for disease symptom development on potatoplants that received the Endura fungicide and the water controltreatment. Foliar application of formulated Bt.4Q7Flg22 (SEQ ID NO: 226)provided as a pre-treatment to potato plants resulted in a disease scoreof 1.83 as compared to plants that received the water control (diseasescore=2.83). Plants that received pre-treatment with the Endurafungicide had the least disease symptoms with a disease score of 0.50 (pvalue=0.0045) compared to plants treated with the water control. Theformulated Bt.4Q7Flg22 polypeptide pre-treatment resulted in plants withan average disease score similar to the enterokinase activatedBt.4Q7Flg22 (H101 EK-activated) provided in a filtrate (fermentationproduct)—both had disease scores of 1.83. The non-activated EK orGST-EK-Bt.4Q7Flg22 or non-cleaved Flg22 filtrate (H101 non-activated)provided to plants had a score of 2.33 and was not significantlydifferent from the disease score of plants that were treated with thewater control (p value=0.4835). However, potato plants that received thepre-treatment with the EK-activated Bt.4Q7Flg22 filtrate resulted in anincreased average stem fresh and dry weight per plant compared acrossall treatments with approximately a 20 g increase in stem fresh weightand an almost 6 g increase in stem dry weight per plant compared toplants that received the water control pre-treatment. Plants thatreceived the formulated Bt.4Q7Flg22 polypeptide all had increased stemfresh and dry weight as measured on a per plant basis compared to plantsthat received the water only control application.

Example 51: Treatment of Candidatus liberibacter asiaticus Infectionwith Flg22 and Anti-Microbial Polypeptides

Bt.4Q7Flg22 formulations were applied by trunk injection treatments toboth Valencia orange (Citrus sinensis) and Ruby Red grapefruit (Citrus xparadisi) trees. The study was conducted at a commercial grove orchardlocated in central Florida (Okeechobee county). Injection treatmentusing the Bt.4Q7Flg22 polypeptide (SEQ ID NO: 226) provided using a 1×Low Rate (0.55 micromoles peptide; 0.138 μM estimated concentration inphloem) and a 10× High Rate (5.5 micromoles peptide; 1.38 μM estimatedconcentration in phloem) was compared to the non-treated control trees.The injection treatments were set up using a randomized complete blockdesign with 10 grapefruit trees (4 years old) per treatment. Theinjections were provided in April (2017) at first flush, a stage ingrowth from the emergence of leaves until they expand to full size.Injection of grapefruit trees were conducted using a low-pressureinjection device, BRANDTENTREE (BRANDT). Leaves from each of thegrapefruit trees were sampled at the time of injection (Day 0), 21 and56 days post injection. A total of six leaf samples per tree wereselected to represent the population of leaves on the tree in terms ofleaf age, location, and presence of visual symptoms. Each midrib wasseparated from the leaf blade and immediately chopped into very smallpieces with a new sterile razor blade. Leaf samples from each tree werethen placed in an individual tube that was subsequently stored in afreezer at −80° C. until further processing. DNA extraction andreal-time polymerase chain reaction or quantitative PCR (qPCR) analysison these leaves was performed at Southern Gardens Citrus (Clewiston,Fla.).

The presences of the CLas bacterial titers in the HLB infected citrustrees can be determined with quantitative real-time polymerase chainreaction (qPCR) methods using specific primers to confirm the presenceof the disease (Li, W. B., Hartung, J. S. and Levy, L. 2008 “Optimizedquantification of unculturable ‘Candidatus liberibacter spp.’ In hostplants using real-time PCR”, Plant Disease 92: 854-861). DNA extractionand quantitative PCR (qPCR) analysis on these leaves was performed atSouthern Gardens Citrus (Clewiston, Fla.) using HLB primer set targetingthe 16S DNA of C. liberibacter bacteria 5′» 3′ (forward): HLB asTCGAGCGCGTATGCAATACG (SEQ ID NO: 773); (reverse) HLBr:GCGTTATCCCGTAGAAAAAGGTAG (SEQ ID NO: 774); HLBpc (probe):AGACGGFTGAGTAACGCG (SEQ ID NO: 775) labeled with an intercalatingfluorescent reporter dye]. Forty cycles of qPCR were conducted and thefluorescent signal which is proportional to the amount of dsDNA insolution was measured. The qPCR analysis allows for the detection of theCLas bacteria in citrus tissue. The cycle threshold (Ct) values from theqPCR analysis were obtained per each treatment. The Ct measurement isequivalent to the number of PCR cycles required to produce a relativethreshold level. As in common practice within the field of molecularbiology, the change in Ct value is reported to indicate the relativequantity of CLas DNA either in treated vs untreated samples or intreated samples at one time point vs another time. The higher the Ctvalue, the greater or more effective the treatment effect, which isindicated by the reduction/elimination of CLas bacteria from the tree. Apercentage reduction in bacterial load can be computed as:

% reduction in sample overtime=(1−2^([Ct(inital time)−Ct(later time)]))*100%

or

% reduction in treated vs. controlsample=(1−2^([Ct(control sample)−Ct(treated sample)]))*100%

The results from the grapefruit trial are shown in FIG. 9. The averagevalues from the Ct comparisons (n=10 trees per treatment) obtained fromthe qPCR analysis from the T0 timepoint (day of injection), the T21 andthe T56 timepoints (21 and 56 days post injection) are reported with thestandard error from the mean Ct values in FIG. 9 (T0=dark grey bars;T21=white bars; T56=light gray bars; average Ct values marked with an“x”). Any outlier values are indicated by the small circles locatedoutside the standard error bars for each treatment. The control orgrapefruit trees that were not injected had the lowest Ct values in arange of Ct near 25 for all treatment timepoints. Leaves sampled fromgrapefruit trees that received injection treatments with the 1× and 10×Bt.4Q7Flg22 polypeptide formulations resulted in slightly higher Ctcounts as compared to leaves from the control trees (FIG. 9). The higherthe Ct. value, the greater the treatment effect for controlling orreducing the infection of the CLas bacteria from spreading. The averageCt value in leaves taken from the T21 sampling was greater than the Ctvalue from the T56 sampling but both were significantly increased overthe non-injected control leaves or leaves from trees that receivedinjections with the Bt.4Q7Flg22 polypeptide formulations (FIG. 9,average Ct values marked with “x”).

In another study using Valencia orange (Citrus sinensis) also conductedat the commercial grove orchard located in central Florida (Okeechobeecounty). Injection treatments using formulations of Bt.4QFlg22 (SEQ IDNO: 226) were compared to antimicrobial polypeptides known as thionins.Thionin injection was provided as a mixture of thionin polypeptides (SEQID NOs: 651, 652 and 653) which are characterized as “un-tagged” orwithout a phloem localization sequence. In addition to the un-taggedthionin mixture, a “tagged” thionin polypeptide that comprised a phloemlocalization sequence (SEQ ID NO: 650) was used as a comparativeinjection treatment. The phloem targeted or “tagged” version was used totarget the thionin specifically to the phloem where CLas bacteria resideand multiply. The injection treatments were applied to orange treesusing a randomized complete block design with a total of 8 orange trees(8 years old) per treatment for the untreated control and Bt.4QFlg22treatments, and a total of 5 orange trees per treatment for the thionintreatments. The injections were provided in April (2017) at first flush,a stage in growth from the emergence of leaves until they expand to fullsize. Injection of the orange trees were conducted using a low-pressureinjection device, BRANDTENTREE (BRANDT). The Bt.4Q7Flg22 polypeptide 1×(0.138 μM) and a 10× (1.37 μM) concentrations, the “untagged” and the“tagged” thionin polypeptides were all compared to trees that receivedno injection treatment (control). Leaves from the orange trees weresampled per each treatment at the time of injection (Day 0) and at T56,or 56 days post injection.

A total of six leaf samples per tree, were selected to represent thepopulation of leaves on the tree in terms of leaf age, location, andpresence of visual symptoms. Each midrib was separated from the leafblade and immediately chopped into very small pieces with a new sterilerazor blade. Leaf samples from each tree were then placed in anindividual tube that was subsequently stored in a freezer at −80° C.until further processing. DNA extraction and real-time polymerase chainreaction or quantitative PCR (qPCR) analysis on these leaves wasperformed at Southern Gardens Citrus (Clewiston, Fla.) using the methodsas described above for performing Ct analysis.

Results from the Valencia orange trial are shown in FIG. 10 (T0=darkgrey bars; T56=white bars). Leaf tissues from the control orange treeshad the lowest Ct values in a range of Ct near 25-30 for treatmenttimepoints T0 and T56 indicating that titer levels of the CLas bacteriadid not change in these trees. Both of the thionin treatments “untagged”and “tagged” had higher average Ct values in leaves taken from the T56sampling as compared to the average Ct values from T56 leaves sampledfrom the water-injected controls (FIG. 10; average Ct values marked with“x”). Any outlier values are indicated by the small circles locatedoutside the standard error bars for each treatment. Leaves sampled fromtrees that received the phloem targeted thionin “tagged” treatment had ahigher average Ct value at T56 compared to leaves from trees thatreceived non-targeted or “un-tagged” thionin treatment. Leaves sampledfrom orange trees that received injection treatments with the 1× and 10×Bt.4Q7Flg22 polypeptide formulations resulted in significantly higher Ctcounts from the T0 to T56 timepoints shown by the average increase in Ctat T56 compared to T0 (FIG. 10; average Ct values marked with “x”).Leaves from trees injected with both Bt.4Q7Flg22 polypeptideformulations (1× and 10×) also had significantly higher Ct valuescompared to leaves samples from the control trees. The Bt.4Q7Flg22 areeffective treatments for controlling or reducing the titer levels of theCLas bacteria in the infected orange trees (FIG. 9).

The Bt.4Q7Flg22 polypeptides provided as injection treatments usingfinal concentrations at the 1× (0.138 μM) and 10× (1.38 μM) were botheffective in reducing CLas titer levels in the leaf tissue sampled 8weeks post injection. The higher concentration of the Bt.4Q7Flg22polypeptide 10× (1.38 μM) however was even more effective resulting in a37% reduction (Trial 1) and a 43% reduction (Trial 2) in CLas titerlevels.

TABLE 84 Treatment effectiveness of Bt.4Q7Flg22 on reducing CLasbacterial titer levels 8 weeks post injection treatment on citrus(Valencia orange and Ruby Red Grapefruit) Percentage Reduction in CLastiter Injection Treatment Normalized to the Control Concentration Trial1 Trial 2 1× Bt.4Q7Flg22 33% 21% (SEQ ID NO: 226) 0.138 μM estimatedconcentration in tree vasculature 10× Bt.4Q7Flg22 37% 43% (SEQ ID NO:226) 1.38 μM estimated concentration in tree vasculature

Previous results indicate that Bt.4Q7Flg22 (SEQ ID NO: 226) promotesplant growth throughout periods of disease (Example 50). To assess for apotential plant growth benefit to injecting HLB-infected ‘Valencia’Orange and ‘Ruby Red’ Grapefruit trees with Bt.4Q7Flg22 (SEQ ID NO:226), current year growth was measured in May 2018 for the same treesthat were injected with Bt.4Q7Flg22 in April 2017 and assessed for CLasbacterial titer at the commercial grove orchard located in centralFlorida (Okeechobee county). Each tree was visually assessed for regionsof current season growth with green color to the branches, as comparedto old growth branches that are more woody in appearance with a darkgreenish-brown to brown hue. Three representative branches with newgrowth were selected per tree, and the distance in inches from the startof green growth (oldest node) to the tip of the youngest node wasmeasured with a flexible measuring tape. Data was collected for treesinjected with 1× and 10× Bt.4Q7Flg22 (SEQ ID NO: 226) as well as theuntreated control, with 8 trees per treatment for the ‘Valencia’ orangetrial and 9-10 trees per treatment for the ‘Ruby Red’ Grapefruit trial(n=24-30 measurements per treatment). Only one tree in the ‘Ruby Red’Grapefruit trial was lost from the original trial (1× Bt.4Q7Flg22treatment group), presumably due to hurricane-strength wind damage inSeptember 2017. For each trial, the average new growth length (inches)was calculated and normalized to the untreated control (Table 85).

TABLE 85 Bt.4Q7Flg22 trunk injection increases new branch growth in‘Valencia’ orange and ‘Ruby Red’ grapefruit Average Flush Length Flushlength Trial Treatment (inches) (% of control) Valencia Control 7.23100% Orange - 1× Bt.4Q7Flg22 13.33 184% Injected (SEQ ID NO: 226) April2017, 0.138 μM estimated Measured May concentration in tree 2018vasculature 10× Bt.4Q7Flg22 12.29 170% (SEQ ID NO: 226) 1.38 μMestimated concentration in tree vasculature Red Control 8.05 100%Grapefruit - 1× Bt.4Q7Flg22 10.83 135% Injected (SEQ ID NO: 226) April2017, 0.138 μM estimated Measured May concentration in tree 2018vasculature 10× Bt.4Q7Flg22 9.13 113% (SEQ ID NO: 226) 1.38 μM estimatedconcentration in tree vasculature

These results demonstrate the ability of Flg22 compositions, whichdisplayed reduced CLas bacterial titer compared to untreated plants(FIG. 9 and FIG. 10), to also enhance the growth of sweet orange andgrapefruit trees (Table 85). Enhanced branch growth serves as anindicator of enhanced fruit yield as more leaves are produced to sustainfruit growth throughout the season. In comparison to the untreatedcontrol, orange and grapefruit trees receiving the 1× Bt.4Q7Flg22injection in April 2017 had on average 6.1 more inches (+85%) or 2.8more inches (+35%) of new branch growth, respectively. The 10× injectiondose of Bt.4Q7Flg22 was also effective at increasing growth, with 5.1more inches (70%) and +1.1 more inches (+13%) of new branch growth inorange and grapefruit trees, respectively. As the 10× Bt.4Q7Flg22injection did not perform better than the 1× injection for enhancinggrowth in 2018, and bacterial titer reductions were similar in 2017. The1× Bt.4Q7Flg22 injection provides a sufficient response in the plant.Importantly, growth measurement indicated that no phytotoxicity occurredafter Flg22 trunk injection at either the 1× or 10× rate.

As these plants were not 100% cleared of disease-causing bacteria, theseresults also demonstrate the ability of the plants injected withBt.4Q7Flg22 to continue to grow despite the presence of HLB-causingbacteria. Provided that CLas strains with antibiotic resistance arepredicted to emerge and become an additional hurdle for HLB-control,Flg22 injection represents a desirable alternative to antibiotictreatments for ameliorating plant growth and reducing bacterial titer.The trees receiving the Flg22 injections in this example were maintainedwith a standard commercial citrus treatment program, which furtherdemonstrates the ability to add Flg22 citrus injections to standardgrower practices.

Example 52: Foliar and Trunk Injection of Flg22 Applied Alone or inCombination with Antimicrobial or Plant-Health Promoting CompoundsIncrease New Shoot Growth in Orange Trees

In subsequent trials in April (2018), Flg22 formulations were applied bytrunk injection treatments or foliar spray at two independent trialsites. Trials were designed to 1) test Flg22 polypeptide variantsproduced synthetically and by fermentation, 2) compare the efficacy ofthe Flg22 variant previously used for citrus injection trials in 2017,Bt.4Q7Flg22 (SEQ ID NO: 226), versus Syn01Bt.4Q7Flg22 (SEQ ID NO: 571)which was effective as both a foliar and seed treatment for increasingyield in row crops, 3) compare Flg22 application methods, namely trunkinjection versus foliar spray to the canopy, and 4) test combinatorialtreatments between Flg22 peptides and oxytetracycline injection,L-cysteine, and Benzo (1,2,3) thiadiazole-7-carbothioic acid-S-methylester (also known as BTH) as the commercially available formulationACTIGARD WG. L-cysteine is an essential, proteinogenic amino acid; andBTH is a salicylic acid analog with increased stability that is usedagriculturally as an activator of plant immune responses and is approvedfor application to citrus trees as root drench or irrigation treatmentto prevent citrus canker caused by Xanthomonas axonopodis pv citri.

In March 2018, trees were treated at two separate sites. Three-year oldHamlin orange trees (Citrus sinensis) were treated at a commerical groveorchard located in central Florida (Okeechobee County). A similar trialwas conducted in a commercial grove of 6-year old Vernia orange trees onSwingle rootstock at Lake Wales, Fla. (Polk County). Treatments wereapplied as listed in Table 85 below using a low-pressure injectiondevice, BRANDT ENTREE (BRANDT) for trunk injection or a CO2-pressurizedbackpack sprayer that produced a fine mist for foliar spray. Trunkinjections were as described in Example 51. Foliar compositions ofBt.4Q7Flg22 were diluted in water with a non-ionic surfactant (PrecisionLabs NIS90:10; 0.1% v/v of spray tank volume) and evenly applied to thecanopy of the tree at a spray rate of 3 Liters per tree. Blocks of treesreceiving a foliar treatment were spaced in the trial area with a gap(skipped tree) in between treatment blocks to avoid drift of treatmentinto neighboring treatment blocks. Treatments were applied during theearly morning or late evening during a period of low wind (<5 mph), andconditions were such all spray treatments dried on leaves within aperiod of 4 hours. Combination treatments described in Table 86 wereeither co-injected in the same BRANDT ENTREE bottle (Citrus Composition7, Citrus Composition 8) or applied separately as an oxytetracyclineinjection followed by a Bt.4Q7Flg22-Syn01 foliar treatment on the sameday (Citrus Composition 11, Citrus Composition 12). For all treatments,10 trees were used per treatment, separated into two replicated blocksof five trees each. Citrus compositions 1-8 were applied at both theOkeechobee and Polk County groves, while Citrus Compositions 9-12 wereapplied at Okeechobee grove alone.

TABLE 86 Treatment compositions tested for ameliorating the effects ofHLB in orange trees Treatment Application Composition Formulation MethodUse Rate Citrus Bt.4Q7Flg22 Trunk 2.75 mL/tree Composition 1 (SEQ ID NO:226) Injection (estimated 0.138 μM 100 μM in plant vasculature) 10 mMSodium Phosphate Buffer, pH 5.7 Citrus Bt.4Q7Flg22-Syn01 Trunk 2.75mL/tree Composition 2 (SEQ ID NO: 571) Injection (estimated 0.138 μM 100μM in plant vasculature) 10 mM Sodium Phosphate Buffer, pH 5.7 CitrusBt.4Q7Flg22 Trunk 80 mL/tree Composition 3 (SEQ ID NO: 226) Injection(fermentation brothfiltrate) With (+) Enterokinase 0.8 U/mL CitrusBt.4Q7Flg22-Syn01 Trunk 80 mL/tree Composition 4 (SEQ ID NO: 571)Injection (fermentation broth filtrate) With (+) Enterokinase 0.8 U/mLCitrus Bt.4Q7Flg22-Syn01 Foliar 3.0 mL/tree in a spray carrierComposition 5 (SEQ ID NO: 571) Spray volume of 3 L water + 0.1% 100 μMv/v Precision Labs NIS90:10 10 mM Sodium Phosphate Buffer, pH 5.7 CitrusBt.4Q7Flg22-Syn01 Foliar 12.0 mL/tree in a spray carrier Composition 6(SEQ ID NO: 571) Spray volume of 3 L water + 0.1% 100 μM v/v PrecisionLabs NIS90:10 10 mM Sodium Phosphate Buffer, pH 5.7 Citrus Part A Trunk2.75 mL/tree Composition 7 Bt.4Q7Flg22-Syn01 Injection (estimated 0.138μM (SEQ ID NO: 571) in plant vasculature) 100 μM 10 mM Sodium PhosphateBuffer, pH 5.7 Part B Trunk 20 mL/tree ACTIGARDWG Injection (1 g pertree) (Active Ingredient: 50% Acibenzolar-S-methyl: Benzo (1,2,3)thiadiazole- 7-carbothioic acid-S-methyl ester; BTH) (50 mg/mL solutionin water) Citrus Part A Trunk 2.75 mL/tree Composition 8Bt.4Q7Flg22-Syn01 Injection (estimated 0.138 μM (SEQ ID NO: 571) inplant vasculature) 100 μM 10 mM Sodium Phosphate Buffer, pH 5.7 Part BTrunk 20 mL/tree L-Cysteine Injection (60 mg per tree) (3 mg/mL solutionin water) Citrus Part A Foliar 3.0 mL/tree in a spray carrierComposition 10 Bt.4Q7Flg22-Syn01 Spray volume of 3 L water + 0.1% (SEQID NO: 571) v/v Precision Labs NIS90:10 100 μM 10 mM Sodium PhosphateBuffer, pH 5.7 Part B Trunk 20 mL/tree Oxytetracycline-HCl Injection(0.45 g per tree) (22.5 mg/mL solution in water) Citrus Part A Foliar12.0 mL/tree in a spray carrier Composition 11 Bt.4Q7Flg22-Syn01 Sprayvolume of 3 L water + 0.1% (SEQ ID NO: 571) v/v Precision Labs NIS90:10100 μM 10 mM Sodium Phosphate Buffer, pH 5.7 Part B Trunk 20 mL/treeOxytetracycline-HCl Injection (0.45 g per tree) (22.5 mg/mL solution inwater)

To assess for a potential plant growth benefit to injecting or sprayingHLB-infected orange trees with different formulations of Flg22polypeptides alone or in combination with antimicrobial or plant-healthpromoting compounds, new flush length was measured in May 2018 for treesthat were treated in March 2018 at commercial groves in Okeechobeecounty, FL and Polk county, FL. At the time of treating plants at bothlocations (March 2018), trees exhibited darker green leaves with 2018season fruit beginning to develop. In the two-month interval betweentreatment (March 2018) and the time of tree measurement (May 2018),trees entered a period of spring flush with new growth visible as verylight green, flexible branches with similarly light green leaves. Eachtree was assessed for new flush, and three representative branches withnew growth were selected per tree. The distance in inches from the startof light green growth (oldest node) to the tip of the youngest node wasmeasured with a flexible measuring tape. Data was collected for 10 treesper treatment including the untreated control, for a total of 30measurements per treatment. Represented in Table 86 is the average flushlength (inches) for each treatment across the two grove sites inOkeechobee and Polk counties, with growth normalized to the untreatedcontrol.

TABLE 87 Flg22 variants applied as either a trunk injection or foliarspray increase new branch growth in ‘Hamlin’ and ‘Vernia’ orange treesAverage Flush Flush length Treatment Length (inches) (% of control)Untreated Control 3.08 100% Citrus Composition 1 3.84 125% Bt.4Q7Flg22(SEQ ID NO: 226) 2.75 mL/tree injection Citrus Composition 2 4.48 146%Bt.4Q7Flg22-Syn01 (SEQ ID NO: 571) 2.75 mL/tree injection CitrusComposition 3 5.18 169% Bt.4Q7Flg22 (SEQ ID NO: 226) Enterokinase (EK) -activated filtrate 80 mL/tree injection Citrus Composition 4 3.28 107%Bt.4Q7Flg22-Syn01 (SEQ ID NO: 571) Enterokinase (EK)- activated filtrate80 mL/tree injection Citrus Composition 5 3.66 119% Bt.4Q7Flg22-Syn01(SEQ ID NO: 571) 1× foliar spray Citrus Composition 6 3.76 122%Bt.4Q7Flg22-Syn01 (SEQ ID NO: 571) 4× foliar spray

Growth measurements of ‘Hamlin’ and ‘Vernia’ new shoots, taken twomonths after either trunk injection or foliar spray application of Flg22variants, indicated that Bt.4Q7Flg22 (SEQ ID NO: 226) and Syn01Flg22(SEQ ID NO: 571) are both effective at promoting greater growth than theuntreated control. On average, untreated control shoots were 3.08 inchesin length, while Bt.4Q7Flg22-injected trees had 25% longer shoots (3.84inches) and Syn01Flg22 injected trees 146% longer shoots (4.48 inches).Bt.4Q7Flg22 and Syn01Flg22 produced through fermentation methodsdescribed in Example 49 were also effective at increasing shoot growthwhen injected into the trunk at a rate of 80 mL/tree. Citrus composition3 containing Bt.4Q7Flg22 produced by fermentation of strain H101 andtreated with 0.8 U/mL Enterokinase (New England Biolabs; Product CodeP8070) was the most effective, with shoots measuring on average 169%(5.18 inches) longer than the untreated control.

Foliar application of Flg22 variants, which is effective for promotinggrowth of kiwi, soy, lentils, and potatoes under disease pressure werealso tested for the ability to promote growth of HLB-infected orangetrees. Table 88 shows that Citrus Compositions 5 and 6 comprised of a 1×or 4× dose of Syn01Bt.4Q7Flg22 (SEQ ID NO: 571), respectively, are alsoeffective at promoting new shoot growth in orange trees. The 1× and 4×doses were similarly effective, with the 1× foliar rate measuring 119%longer shoots than the control, and the 4× rate measuring 122% longershoots than the control. These results show that a foliar application ofFlg22 polypeptide can be used as part of a standard program of care ofcitrus grove trees.

TABLE 88 Injection of Bt.4Q7Flg22-Syn01 in combination with plant-healthpromoting compounds increases new branch growth in ‘Hamlin’ and ‘Vernia’orange trees Average Flush Flush length Treatment Length (inches) (% ofcontrol) Untreated Control 3.08 100% Citrus Composition 2 4.48 146%Bt.4Q7Flg22-Syn01 (SEQ ID NO: 571) 2.75 mL/tree injection CitrusComposition 7 3.44 112% Bt.4Q7Flg22-Syn01 (SEQ ID NO: 571) 2.75 mL/treeinjection + BTH (ACTIGARD WG; 1 g/tree injection) Citrus Composition 85.87 191% Bt.4Q7Flg22-Syn01 (SEQ ID NO: 571) 2.75 mL/tree injection +L-Cysteine (60 mg/tree injection)

Next, the combination of Syn01Bt.4Q7Flg22 (SEQ ID NO: 571) with BTH(ACTIGARD WG) or L-cysteine was investigated at both the Melvinlocations and Lake Wales groves. Both combination treatments in Table 88showed greater new flush length in comparison to the untreated control,showing that Flg22 polypeptides can be used in combination with aminoacids, plant hormones, or plant hormone-mimics to improve citrus treehealth.

TABLE 89 Foliar spray application of Bt.4Q7Flg22-Syn01 in combinationwith oxytetracycline injection increases new branch growth in 3-year old‘Hamlin’ orange trees Treatment Average Flush Flush Length (see table85) Length (inches) (% of control) Untreated Control 1.67 100% CitrusComposition 9 3.80 228% Oxytetracycline-HCl (0.45 g/tree) + Syn01Bt.4Q7Flg22 (SEQ ID NO: 571) 1× foliar spray Citrus Composition 10 3.32199% Oxytetracycline-HCl (0.45 g/tree) + Syn01 Bt.4Q7Flg22 (SEQ ID NO:571) 4× foliar spray

In a separate trial, the combination of Syn01Flg22 (SEQ ID NO: 571) andoxytetracycline treatments were observed. On the same day that treeswere injected with oxytetracycline, groups of 10 trees were also sprayedwith a foliar application of Syn01Flg22 at a 1× rate or 10× rate. Theseresults show that the antibiotic and polypeptide treatments arecompatible and that no phytoxicity was observed due to the dualtreatment. A standard program could be envisioned where groweralternated tree injections with foliar treatments for enhanced controlof HLB symptoms and for reducing CLas titer.

Example 53: Disease Protection Using Bt.4Q7Flg22 and Gm.RHPP FoliarApplications on Soybean Plants to Protect from Diseases Caused byPhakopsora pachyrhizi and Cercospora kikuchii

Table 90 describes the compositions and corresponding use rates testedin the following example.

TABLE 90 Bt.4Q7Flg22 and Gm.RHPP foliar applications on soy protectplants from Phakopsora pachyrhizi and Cercospora kikuchii ApplicationUse Rate Fluid ounce/acre (Fl. oz/Ac) Milliliters/hectare CompositionFoliar Formulation (mL/Ha) Composition 12 FOX Fungicide 5.48 Fl. oz/Acor 400 mL/Ha Composition 13 Bt.4Q7Flg22 2.05 Fl. oz/Ac or (SEQ ID NO:226) 150 mL/Ha 16.7 μM 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BCpreservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) Composition14 Bt.4Q7Flg22 4.11 Fl. oz/Ac or (SEQ ID NO: 226) 300 mL/Ha 16.7 μM 1.67mM Sodium Phosphate Buffer, pH 5.7 PROXEL BC preservative: 330.7 μM(BIT); 53.5 μM (CMIT); 26.1 μM (MIT) Composition 15 Gm.RHPP 2.05 Fl.oz/Ac or (SEQ ID NO: 600) 150 mL/Ha 100 μM PROXEL BC preservative: 330.7μM; 50.1 μM (CMIT); 21.71 μM (MIT) Composition 16 Gm.RHPP 4.11 Fl. oz/Acor (SEQ ID NO: 600) 300 mL/Ha 100 μM PROXEL BC preservative: 330.7 μM;50.1 μM (CMIT); 21.71 μM (MIT) Composition 17 FOX Fungicide + 5.48 Fl.oz/Ac or Bt.4Q7Flg22 400 mL/Ha + (SEQ ID NO: 226) 2.05 Fl. oz/Ac or 16.7μM 150 mL/Ha 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BCpreservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) Composition18 FOX Fungicide + 5.48 Fl. oz/Ac or Bt.4Q7Flg22 400 mL/Ha + (SEQ ID NO:226) 4.11 Fl. oz/Ac or 16.7 μM 300 mL/Ha 1.67 mM Sodium PhosphateBuffer, pH 5.7 PROXEL BC preservative: 330.7 μM (BIT); 53.5 μM (CMIT);26.1 μM (MIT) Composition 19 FOX Fungicide + 5.48 Fl. oz/Ac or Gm.RHPP400 mL/Ha + (SEQ ID NO: 600) 2.05 Fl. oz/Ac or 100 μM 150 mL/Ha PROXELBC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) Composition 20FOX Fungicide + 5.48 Fl. oz/Ac or Gm.RHPP 400 mL/Ha + (SEQ ID NO: 600)300 mL/Ha 100 μM PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71μM (MIT) * Foliar compositions contained 0.1% (v/v) PROXEL BCpreservative, an aqueous dispersion of a blend of 330.7 mM1,2-benzisothiazolin (BIT), 53.5 mM5-chloro-2-methyl-4-isolthiazolin-3-one (CMIT), and 26.1 mM2-methyl-4-isothiazolin-3-one (MIT). Foliar compositions were applied atthe indicated rates (Fl. oz/Ac or mL/Ha) in a carrier volume of 150 L/Haor 16 gallons/acre water with 0.5% (v/v) AUREO methylated bean oilsurfactant (Composition 13) or with 0.33% (v/v) Agris Parrafinic mineraloil (stock concentration of 795 g/L or 79.5% (p/v) (Compositions 13-20).

Replicated field trials were conducted across three locations inParaguay (Yatytay, Obligado, and Capitan Miranda) using a foliarapplication comprising a compositions of the Bt.4Q7Flg22 polypeptide andRHPP polypeptide provided with a broad-spectrum fungicide, Fox (16.0%prothioconazole and 13.7% thiofloxystrobin). FOX is a commerciallyavailable foliar fungicide in South America with limited efficacy forpreventative and curative treatment of Asian soybean rust caused byPhakopsora pachyrhizi and Cercospora leaf blight of soybean caused byCercospora kikuchii applied as a foliar spray following therecommendations on the specimen label at a use rate of 5.48 fluid ouncesper acre (Fl. oz/Ac) (400 mL/hectare). Beginning at the R1 stage ofdevelopment, soybean plants received two foliar applications of thecompositions described in Table 90 with an interval of 13-14 daysbetween spray applications. Foliar treatments were applied to a singlesoy variety (which one? Same at all 3 sites) at the three sites, with 4replicated plots (3×10 meters, 30 m2; with minimum of 6 rows pertreatment). Disease assessments for trials that were naturally infectedwere scored for the severity of infection (0-100% of foliage affected)were scored for 10 plants within each plot for both Asian soybean rustcaused by Phakopsora pachyrhizi and Cercospora leaf blight of soybeancaused by Cercospora kikuchii at the R4-R5 stage of soy development(4-15 days after second foliar application) with guidance from Godoy etal (1997; Journal of plant diseases and protection 104:336-345). Percentphytotoxicity (0-100% of foliage affected) was also scored at the R4-R5stage of soy development. Severity of infection and phytotoxicity wereaveraged across all four replicates per site (Total=12 replicates, 3sites with 4 replicates each). Standard deviation for each treatmentbetween the three sites was calculated. Untreated control plants at theYatytay site displayed 99% defoliation at 11 days post-application ofthe second foliar treatment and were scored for defoliation (0-100%defoliated) at this time. Disease severity, phytotoxicity, anddefoliation results are provided in Table 91 as percentages, withstandard deviation in parentheses.

TABLE 91 Incidence of Asian Soybean Rust disease symptoms after foliarapplication of fungicide and polypeptide compositions in ParaguayIncidence of Asian Change in Defoliation Soybean Rust symptoms AsianSoybean (% of foliage) Application Use Rate after 2 foliar Rustsymptoms, after 2 foliar Fluid ounce/acre applications relative toapplications (Fl. oz/Ac) (% of foliage affected); control (%); (Yatytayonly; Foliar Milliliters/hectare N = 12 reps N = 12 reps N = 4 repsFormulation (mL/Ha) per treatment per treatment per treatment) UntreatedControl n/a  35.1% — 99% (±13.8%) FOX Fungicide 5.48 Fl. oz/Ac or  19.4%−15.7% 45% (Composition 12) 400 mL/Ha (±10.3%) (−15.7%) Bt.4Q7Flg22 2.05Fl. oz/Ac or  22.7% −12.4% 70% (SEQ ID NO: 226) 150 mL/Ha (±14.4%) 16.7μM 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BC preservative: 330.7μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition 13) Bt.4Q7Flg224.11 Fl. oz/Ac or  22.1% −13.0% 60% (SEQ ID NO: 226) 300 mL/Ha (±14.6%)16.7 μM 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BC preservative:330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition 14) Gm.RHPP2.05 Fl. oz/Ac or  24.6% −10.5% 96% (SEQ ID NO: 600) 150 mL/Ha (±13.4%)100 μM PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT)(Composition 15) Gm.RHPP 4.11 Fl. oz/Ac or  23.8% −11.3% 70% (SEQ ID NO:600) 300 mL/Ha (±11.5%) 100 μM PROXEL BC preservative: 330.7 μM; 50.1 μM(CMIT); 21.71 μM (MIT) (Composition 16) FOX Fungicide + 5.48 Fl. oz/Acor  9.3% −25.8% 25% Bt.4Q7Flg22 400 mL/Ha +  (±3.9%) (SEQ ID NO: 226)2.05 Fl. oz/Ac or 16.7 μM 150 mL/Ha 1.67 mM Sodium Phosphate Buffer, pH5.7 PROXEL BC preservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM(MIT) (Composition 17) FOX Fungicide + 5.48 Fl. oz/Ac or  8.0% −27.1%25% Bt.4Q7Flg22 400 mL/Ha +  (±5.1%) (SEQ ID NO: 226) 4.11 Fl. oz/Ac or16.7 μM 300 mL/Ha 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BCpreservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition18) FOX Fungicide + 5.48 Fl. oz/Ac or  8.3% −26.8% 25% Gm.RHPP 400mL/Ha +  (±5.8%) (SEQ ID NO: 600) 2.05 Fl. oz/Ac or 100 μM 150 mL/HaPROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT)(Composition 19) FOX Fungicide + 5.48 Fl. oz/Ac or  11.0% −24.1% 25%Gm.RHPP 400 mL/Ha +  (±3.3%) (SEQ ID NO: 600) 300 mL/Ha 100 μM PROXEL BCpreservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 20)

TABLE 92 Incidence of Cercospora leaf blight symptoms after foliarapplication of fungicide and polypeptide compositions in ParaguayIncidence of Cercospora symptoms Application Use Rate after 2 foliarChange in Fluid ounce/acre applications, (% Cercospora symptoms, (Fl.oz/Ac) of foliar affected); relative to control Milliliters/hectare N =12 reps per (%); N = 12 reps Foliar Formulation (mL/Ha) treatment pertreatment Untreated Control n/a 19.3% (±5.1%) — FOX Fungicide 5.48 Fl.oz/Ac or 15.0% (±9.6%) −4.3% (Composition 12) 400 mL/Ha Bt.4Q7Flg22 2.05Fl. oz/Ac or 16.4% (±7.7%) −2.8% (SEQ ID NO: 226) 150 mL/Ha 16.7 μM 1.67mM Sodium Phosphate Buffer, pH 5.7 PROXELBC preservative: 330.7 μM(BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition 13) Bt.4Q7Flg22 4.11Fl. oz/Ac or 15.8% (±6.3%) −3.5% (SEQ ID NO: 226) 300 mL/Ha 16.7 μM 1.67mM Sodium Phosphate Buffer, pH 5.7 PROXELBC preservative: 330.7 μM(BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition 14) Gm.RHPP 2.05 Fl.oz/Ac or 15.9% (±6.9%) −3.3% (SEQ ID NO: 600) 150 mL/Ha 100 μM PROXELBCpreservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 15)Gm.RHPP 4.11 Fl. oz/Ac or 14.8% (±5.3%) −4.5% (SEQ ID NO: 600) 300 mL/Ha100 μM PROXELBC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT)(Composition 16) FOX Fungicide + 5.48 Fl. oz/Ac or 10.6% (±3.9%) −8.7%Bt.4Q7Flg22 400 mL/Ha + (SEQ ID NO: 226) 2.05 Fl. oz/Ac or 16.7 μM 150mL/Ha 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXELBC preservative:330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition 17) FOXFungicide + 5.48 Fl. oz/Ac or 10.6% (±4.4%) −9.2% Bt.4Q7Flg22 400mL/Ha + (SEQ ID NO: 226) 4.11 Fl. oz/Ac or 16.7 μM 300 mL/Ha 1.67 mMSodium Phosphate Buffer, pH 5.7 PROXELBC preservative: 330.7 μM (BIT);53.5 μM (CMIT); 26.1 μM (MIT) (Composition 18) FOX Fungicide + 5.48 Fl.oz/Ac or 11.3% (±4.2%) −8.0% Gm.RHPP 400 mL/Ha + (SEQ ID NO: 600) 2.05Fl. oz/Ac or 100 μM 150 mL/Ha PROXEL BC preservative: 330.7 μM; 50.1 μM(CMIT); 21.71 μM (MIT) (Composition 19) FOX Fungicide + 5.48 Fl. oz/Acor 11.3% (±4.2%) −8.0% Gm.RHPP 400 mL/Ha + (SEQ ID NO: 600) 300 mL/Ha100 μM PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT)(Composition 20)

TABLE 93 Phytotoxicity after foliar application of fungicide andpolypeptide compositions in Paraguay Application Phytotoxicity (% of UseRate foliage affected) Fluid ounce/acre after 2 foliar (Fl. oz/Ac)applications; Milliliters/hectare N = 12 reps per Foliar Formulation(mL/Ha) treatment Untreated Control n/a 0.00% (±0.00%) FOX Fungicide5.48 Fl. oz/Ac or 2.25% (±32.9%) (Composition 12) 400 mL/Ha Bt.4Q7Flg222.05 Fl. oz/Ac or 0.00% (±0.00%) (SEQ ID NO: 226) 150 mL/Ha 16.7 μM 1.67mM Sodium Phosphate Buffer, pH 5.7 PROXEL BC preservative: 330.7 μM(BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition 13) Bt.4Q7Flg22 4.11Fl. oz/Ac or 0.00% (±0.00%) (SEQ ID NO: 226) 300 mL/Ha 16.7 μM 1.67 mMSodium Phosphate Buffer, pH 5.7 PROXEL BC preservative: 330.7 μM (BIT);53.5 μM (CMIT); 26.1 μM (MIT) (Composition 14) Gm.RHPP 2.05 Fl. oz/Ac or0.00% (±0.00%) (SEQ ID NO: 600) 150 mL/Ha 100 μM PROXEL BC preservative:330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 15) Gm.RHPP 4.11Fl. oz/Ac or 0.00% (±0.00%) (SEQ ID NO: 600) 300 mL/Ha 100 μM PROXEL BCpreservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 16)FOX Fungicide + 5.48 Fl. oz/Ac or 2.33% (±0.14%) Bt.4Q7Flg22 400 mL/Ha +(SEQ ID NO: 226) 2.05 Fl. oz/Ac or 16.7 μM 150 mL/Ha 1.67 mM SodiumPhosphate Buffer, pH 5.7 PROXEL BC preservative: 330.7 μM (BIT); 53.5 μM(CMIT); 26.1 μM (MIT) (Composition 17) FOX Fungicide + 5.48 Fl. oz/Ac or2.25% (±0.25%) Bt.4Q7Flg22 400 mL/Ha + (SEQ ID NO: 226) 4.11 Fl. oz/Acor 16.7 μM 300 mL/Ha 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BCpreservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition18) FOX Fungicide + 5.48 Fl. oz/Ac or 2.42% (±0.29%) Gm.RHPP 400 mL/Ha +(SEQ ID NO: 600) 2.05 Fl. oz/Ac or 100 μM 150 mL/Ha PROXEL BCpreservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 19)FOX Fungicide + 5.48 Fl. oz/Ac or 2.17% (±0.29%) Gm.RHPP 400 mL/Ha +(SEQ ID NO: 600) 300 mL/Ha 100 μM PROXEL BC preservative: 330.7 μM; 50.1μM (CMIT); 21.71 μM (MIT) (Composition 20)

Foliar application of Bt.4Q7Flg22 and Gm.RHPP during reproductive phasesof soy development provided increased protection against Asian soybeanrust and Cercospora leaf blight as compared to the untreated control.Foliar applications of Bt.4Q7Flg22-treated plants at 150 and 300 mL/Hadisplayed 12.4-13.0% less Asian soybean rust leaf area damage and2.8-3.5% less Cercospora leaf area damage compared to the untreatedcontrol; and foliar applications of Gm.RHPP-treated plants at 150 and300 mL/Ha displayed 10.5-11.3% less Asian soybean rust leaf area damageand 3.3-4.5% less Cercospora leaf area damage compared to the untreatedcontrol. Combination treatments including either Bt.4Q7Flg22 or RHPPwith FOX fungicide increased protection against Asian Soybean Rust andCercospora relative to the Fox Fungicide treatment alone. At the Yatytaysite, less defoliation was observed at the R7 stage of development dueto severe disease symptoms upon Bt.4Q7Flg22 or Gm.RHPP treatments +/−FOXfungicide. While the untreated control was 99% defoliated at this stage,Bt.4Q7Flg22 treatment at 150 or 300 mL/Ha decreased defoliation to 70 or60% with green leaves still visible, respectively. The Gm.RHPP treatmentat 150 or 300 mL/Ha decreased defoliation to 96% or 70%, respectively.Combination treatment with Bt.4Q7Flg22 or Gm.RHPP treatments with FOXfungicide decreased defoliation to 25% with green leaves visible, whileFox fungicide alone decreased defoliation to only 45% without greenleaves visible. Overall, polypeptide treatments provided increasedprotection over FOX Fungicide alone for control of Asian soybean rustand Cercospora leaf blight. No phytotoxicity was observed for anypolypeptide application alone, and combination of either polypeptidewith FOX fungicide neither significantly increased or decreasedphytotoxicity relative to the FOX Fungicide alone (Table 93).

Example 54. Flg22-PSA Foliar Application on Kiwi Protects Plants fromPseudomonas syringae pv. Actinidiae (PSA-V)

Pseudomonas syringae pv. actinidiae (PSA) is a devastating plantpathogen causing bacterial canker of both green- (Actinidiae deliciosa)and yellow-flesh (Actinidiae chinesis) kiwi plants throughout zones ofkiwi production, causing severe harvest loss in New Zealand, China, andItaly. In New Zealand alone, cumulative revenue losses to the mostdevastating biovar PSA-V are predicted to approach $740 million NewZealand leaves Dollars (NZD) by 2025 (Agribusiness and EconomicsResearch Institute of Lincoln University “The Costs of Psa-V to the NewZealand Kiwifruit Industry and the Wider Community”; May 2012). PSA-Vcolonizes the outer and inner surfaces of the kiwi plant and can spreadthrough the xylem and phloem tissues. Disease symptoms of PSA-V on kiwiinclude bacterial leaf spot, bacterial canker of the trunk, redexudates, blossom rot, discoloration of twigs, and ultimately dieback ofkiwi vines. The standard method of control for PSA-V currently employsfrequent foliar applications of metallic copper to kiwi vines which ispredicted to lead to the selection of copper-resistant form of thepathogen and loss of disease control. Novel methods of control areurgently needed.

To test the sensitivity of kiwi leaves to 22-amino acid fragments offlagellin, 1 mm slices were cut through Actinidiae deliciosa Kiwi‘Hayward’ leaf petioles and floated in 150 μL of water in a 96-wellplate, with one slice per well. Flg22 polypeptides in Table 94 wereprepared for the assay by re-suspending lyophilized polypeptide indeionized water to a concentration of 10 mM; peptides were then seriallydiluted to 10 μM in 100 mM sodium phosphate (pH 7.8-8.0) buffer with0.1% Tween-20. Water was removed from kiwi leaf petiole samples after 20hours and replaced with 100 μL of an elicitation solution containing 100nM peptide (diluted from 10 μM stock), 34 μg/mL luminol, and 20 μg/mLhorseradish peroxidase in deionized water. Recognition of the Flg22polypeptide by the plant tissue resulted in activation of immunesignaling and the production of apoplastic reactive oxygen species(ROS). In the presence of ROS (H₂O₂), horseradish peroxidase catalyzedthe oxidation of luminol and production of visible light. Relative lightunits (RLUs) were recorded with a SpectraMax L luminometer (0.5 sintegration; 2.0 min intervals) over a time course of 40 minutes. In twoindependent experiments, a total of 6 kiwi leaf petiole samples weretreated with each Flg22 polypeptide in Table 94. The average total RLUand standard error of the means (SEM) was calculated for each treatment.A two-tailed T-test was used to determine significance at the 90%confidence level (P<0.1) between treatments. Relative ROS production wasdetermined for each polypeptide in comparison to total RLUs for the 100nM Bt.4Q7Flg22 control.

TABLE 94 Kiwi leaf petioles are most sensitive to Flg22-PSA AverageTotal Relative Light P-value Units (RLUs); compared to ROS productionSEM in 100 nM relative to Treatment parentheses Bt.4Q7Flg22 Bt4Q7Flg22(%) 100 nM Bt.4Q7Flg22- 47,457 n/a 100% (SEQ ID NO: 226) (±12,900) 100nM Syn01Flg22 81,848 p = 0.286 172% (SEQ ID: 571) (±27,631) 100 nMFlg22-PSA 124,550   p = 0.058* 262% (SEQ ID: 540) (±33,555) *Significantdifference at the 90% confidence level

Across two independent experiments Kiwi ‘Hayward’ leaf petioles weresignificantly more sensitive to Flg22 derived from Pseudomonas syringaepv. actinidiae (Flg22-PSA; SEQ ID NO:540) in comparison to Flg22 derivedfrom Bacillus thuringiensis strain 4Q7 (Bt.4Q7Flg22; SEQ ID NO: 226).While ROS production was increased in kiwi leaf petioles in response tothe synthetic Syn01Flg22 (SEQ ID NO: 571) in comparison to Bt.4Q7 Flg22(SEQ ID NO: 226) the difference was not significant. Based on theseresults, Flg22-PSA (SEQ ID NO: 540) was formulated as indicated at 100nM final concentration (Table 94) for disease prevention trials inpotted Kiwi ‘Hayward’ plants in New Zealand.

TABLE 95 Treatments applied to potted kiwi trial Product dilution forspray application Milliliters product/Liter water (mL/L) or Gramsproduct/Liter Composition Foliar Formulation water (g/L) Composition 21ChampION++ ™ 0.9 g ChampION++ ™/ (46.1% Copper Hydroxide; L water 30%metallic copper equivalent) Composition 22 Flg22-PSA 4 mL/L water (SEQID NO: 540) 100 μM 10 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BCpreservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT)

Foliar compositions contained 0.1% (v/v) PROXEL BC preservative, anaqueous dispersion of a blend of 330.7 mM 1,2-benzisothiazolin (BIT),53.5 mM 5-chloro-2-methyl-4-isolthiazolin-3-one (CMIT), and 26.1 mM2-methyl-4-isothiazolin-3-one (MIT). Foliar compositions were diluted tothe indicated concentrations in water (g/L water or mL/L water) with0.05% (v/v) Contact XceI™ non-ionic surfactant. The diluted productswere applied in fine droplets with a pressurized backpack sprayer to theentire canopy of each plant, until thoroughly covered.

To assess the efficacy of Flg22-PSA (SEQ ID NO: 540) for control ofPseudomonas syringae pv. actinidiae (PSA-V), a potted kiwi disease trialwas conducted in the Bay of Plenty area of New Zealand by HortEvaluationLtd in collaboration with NuFarm Limited. PSA-V symptom-free potted kiwiActinidiae deliciosa ‘Hayward’ plants were evenly distributed betweenthe 6 treatment groups, with 12 potted plants per group. One day priorto inoculation with PSA-V, potted plants were treated with ChampION++™,the industry standard for PSA-V control, or formulated Flg22-PSAaccording to the application rates in Table 96 (Treatment groups 3,4) ata plant nursery in Te Puka, New Zealand. After 24 hours, all plantsexcept for the uninfected controls were sprayed with 1×108 cfu/mL PSA-Vinoculum using a 5 L hand-held pressurized sprayer aimed at theunderside of leaves until thoroughly covered. The uninfected control wassprayed with water alone. Potted plants were then transported toPukehina and placed in an area with overhead misting for 48 hours tomimic environmental conditions for PSA-V infection, with uninfectedcontrol plants separated from infected plants. After 48 hours, a subsetof plants was then removed from the misting area and allowed to brieflydry. After the final treatments, all plants were moved to their finaloutdoor trial site, randomized positions in Pukehina. Average dailytemperature at the trial site was 20.75° C. with a total rainfall of 277mm over 34 days. Additionally, each plant was watered twice a day fortwo hours at a time by drip irrigation. Environmental conditions werefavorable for progression of PSA-V disease symptoms. Plants werevisually monitored throughout the trial period for PSA-V diseaseassessments, with the same assessor recording the % of leaf area coveredin spots at 6 days after inoculation (6 DAI), 16 DAI, 23 DAI and 29 DAI.Additionally, each plant was assessed for treatment phytotoxicityeffects at 29 DAI on a scale of 0-10, with 0=no leaf phytotoxicity and10=very severe leaf phytotoxicity symptoms. The average disease scoresat 6, 16, 23, and 29 DAI and phytotoxicity score at 29 DAI are reportedin Table 96 for each treatment (n=12 plants per treatment). P-valueswere calculated for each treatment vs. the untreated control.

TABLE 96 Flg22-PSA foliar application reduces PSA-V disease symptoms inkiwi plants Application Foliage Affected (% leaf surface area);Treatment group #/ Rate and p-values vs. untreated control FoliarFormulation Timing 6 DAI 16 DAI 23 DAI 29 DAI Treatment group 1 n/a0.00%  1.66%  7.89% 18.14% Uninfected plants Treatment group 2 n/a15.12%  40.36% 54.64% 67.82% Untreated Control Treatment group 3 0.9g/L; 3.23% 12.48% 16.57% 25.20% ChampION++ ™ One day (p < 0.001) (p <0.001) (p < 0.001) (p < 0.001) (Composition 21) pre- inoculationTreatment group 4 4 mL/L; 7.31% 29.41% 45.97% 61.91% Flg22-PSA One day(p < 0.001) (p = 0.013) (p = 0.085) (p = 0.190) (Composition 22) pre-inoculation

Application of Flg22-PSA significantly reduced PSA-V leaf spot symptoms(P<0.1; 90% confidence interval) at 6, 16 and 23 DAI in comparison tothe untreated control. Combination of Flg22-PSA pre-treatment furtherdecreased the severity of leaf spot compared to Flg22-PSA treatmentalone at all assessment timepoints and prolongs the period ofsignificant protection to 29 DAI (14.3% less leaf spot compared tountreated control; P=0.002). In conclusion, Flg22-PSA can be used bothas a stand-alone treatment and in combination with other treatmentsaimed at restricting pathogen growth. While the industry standardChampION++™ which is the currently used copper containing treatment totreat PSA causes mild leaf phytotoxicity (AVE score=1.6), no significantphytotoxicity was observed for Treatments 3-4 (Table 97). Flg22-PSA canbe used as an alternative to other phytotoxic treatments.

TABLE 97 FLG22-PSA foliar application does not cause leaf phytotoxicityof kiwi plants Average Treatment group #/ Application Rate PhytotoxicityScore Foliar Formulation and Timing (0-10); 29 DAI Treatment group 1 n/a0.0 (±0.0) Uninfected plants Treatment group 2 n/a 0.0 (±0.0) UntreatedControl Treatment group 3 0.9 g/L; 1.6 (±0.9) ChampION++ ™ One day pre-(Composition 21) inoculation Treatment group 4 4 mL/L; 0.1 (±0.3)Flg22-PSA One day pre- (Composition 22) inoculation

Example 55: Polypeptides Derived from Elongation Factor Tu

Elf18 and Elf26 polypeptides derived from the consensus Bacillus cereusElongation Factor-TU (EF-Tu) protein were tested for ability to producea ROS response in corn (hybrid 5828 YX), soy (variety Morsoy), andArabidopsis thaliana. Polypeptides were synthesized by Genscript USA(Piscataway, N.J.) using standard solid-phase synthesis methods andprovided as a lyophilized powder with greater than or equal to 70%purity. Dry powder was re-suspended to a concentration of 10 mM inultrapure water, and then serially diluted in ultrapure water to theconcentrations tested in the ROS assay in Table 98.

For the ROS assay, Arabidopsis leaves were excised from 4-week-oldplants, and using a cork borer 4 mm disks were removed from the leaves.Each disc was cut in half using the edge of a razor blade, and then eachdisc half was floated on 150 μL of water abaxial side touching the waterin a 96-well plate to rest overnight. The next day, the water wasremoved from each well just prior to polypeptide treatment. RLU valuesand relative ROS activity was reported as the average of 4 measurements.ROS activity assays were conducted using the methods as previouslyreported in Example 15). ROS activity results are reported in Table 97below.

TABLE 98 Elf18 and Elf26 Polypeptides from Bacillus cereus EF-Tu  AminoPolypeptide Acid Description Length Sequence N terminus of EF Tu 18Ac-AKAKFERSKPHV (modified) NIGTIG-conh2 Bacillus cereus (SEQ ID NO: 616)N terminus of EF Tu 26 Ac-AKAKFERSKPHV (modified) NIGTIGHVDHGKTT-Bacillus cereus conh2 (SEQ ID NO: 617)

TABLE 99 Comparison of ROS activity of elf18 and elf26 polypeptides inArabidopsis leaf tissue Average RLU value (Fold increase (×) PolypeptideTreatment over mock treatment) Negative control (water) 82896 (1×)  Nterminus of EF Tu (100 nM) 264194 (3.2×) (SEQ ID NO: 616) N terminus ofEF Tu (100 nM) 211383 (2.5×) (SEQ ID NO: 617) Bt.4Q7Flg22 (100 nM)258073 (3.1×) (SEQ ID NO: 226) N terminus of EF Tu (100 nM) 254344(3.1×) (SEQ ID NO: 616) + Bt.4Q7Flg22 (100 nM) (SEQ ID NO: 226) Nterminus of EF Tu (100 nM) 181504 (2.2×) (SEQ ID NO: 617) + Bt.4Q7Flg22(100 nM) (SEQ ID NO:226)

The receptor for EF-Tu polypeptides, EF-Tu Receptor (EFR) was previouslyidentified in the Brassica clade, of which Arabidsopis thaliana is amodel plant. Results in Table 99 indicate that newly identifiedpolypeptides from Bacillus cereus EF-Tu (SEQ ID NO: 616 and SEQ ID NO:617) can be used to elicit a ROS response similar in magnitude toBt.4Q7Flg22 (SEQ ID NO: 226) when each was tested at a 100 nMconcentration. In comparison to the mock-treated control, EF-TuN-terminal polypeptides gave a response that was 3.2- to 2.5-foldincreased, while Bt.4Q7Flg22 was 3.1-fold increased over mock control.These results suggest that 18- and 26- amino acid fragments from theN-terminus of Bacillus cereus can be used similarly to Bt.4Q7Flg22 inthe Brassica crops, including but not limited to kale, cabbage, collardgreens, cauliflower, Brussel sprouts, savoy, kohlrabi and gai Ian, toincrease plant biomass, yield and disease prevention.

Combination treatments of EF-Tu N-terminal peptides (SEQ ID NO: 616 andSEQ ID NO: 617) and Bt.4Q7Flg22 (SEQ ID NO: 226) resulted in similar ROSresponses to the EF-Tu peptides alone, indicating that the combinationof peptides treatments in the field would provide no interference ofactivity; however, due to the shared mechanisms between downstreamsignaling events for EF-Tu and Flg22 peptides, recognized by the EFR andFLS2 receptors respectively, a staggered application of peptidetreatments may provide the greatest growth benefit to the plant.

Example 56: Disease Protection Using Bt.4Q7Flg22 and Gm.RHPP FoliarApplications on Soybean Plants to Protect from Diseases Caused byPhakopsora pachyrhizi and Cercospora kikuchii

Foliar application of Bt.4Q7Flg22 (SEQ ID NO: 226) and Gm.RHPP (SEQ IDNO: 600) during reproductive phases of soy development was previouslyfound to decrease disease symptoms caused by Phakopsora pachyrhizi andCercospora kikuchii infections (Example 53). These plants were taken toyield, and Bt.4Q7Flg22 (SEQ ID NO: 226) and Gm.RHPP (SEQ ID NO: 600)foliar applications were found to increaseyield in comparison to theuntreated control plants in replicated trials in Paraguay where plantswere infected with Asian soybean rust and Cercospora leaf blight. Foliarapplications of Bt.4Q7Flg22 at 150 and 300 mL/Ha increased yield by+342.2 kg/Ha and +427.2 kg/Ha, respectively, in trials where the averageyield for untreated plants was 1266.3 Kg/Ha. The increase in yield for300 mL/Ha foliar application of Bt.4Q7Flg22 (36.1%) was comparable toFOX Fungicide alone (36.6%), demonstrating that Bt.4Q7Flg22 is effectiveas an anti-fungal foliar treatment for both reducing disease symptomsand boosting yield. The relative yield across all three trial sites wasfor plants treated with a combined application of FOX Fungicide andBt.4Q7Flg22 was slightly increased over FOX fungicide or Bt.4Q7Flg22foliar application alone, demonstrating that the treatments arecompatible. Foliar applications of Gm.RHPP at 150 and 300 mL/Ha furtherincreased yield by +294.2 kg/Ha and 506.8 kg/Ha, respectively, incomparison to the untreated control. When applied in combination withFOX Fungicide, Gm.RHPP provided the greatest protection against diseasein the trials as evidenced by increased yield of +517.6 kg/Ha and +539.9kg/Ha for the 150 mL/Ha and 300 mL/Ha application rates of Gm.RHPP,respectively. Foliar application of Gm.RHPP consistently improved planthealth and increased yield, thus Gm.RHPP is an effective treatment forgrowth promotion and fungal disease resistance.

TABLE 100 Soybean yield for replicated field trials infected withPhakopsora pachyrhizi and Cercospora kikuchii where plants were treatedwith Bt.4Q7Flg22 or RHPP Average change in yield in comparison toUntreated Control Application Use Rate [1266.3 Yield relative Fluidounce/acre Kilograms/Hectare to Untreated (Fl. oz/Ac) (Kg/Ha)]; Control(%); Milliliters/hectare N = 12 reps per N = 12 reps per FoliarFormulation (mL/Ha) treatment treatment Untreated Control n/a —   100%FOX Fungicide 5.48 Fl. oz/Ac or +445.6 kg/Ha 136.6% (Composition 12) 400mL/Ha Bt.4Q7Flg22 2.05 Fl. oz/Ac or +342.2 kg/Ha 130.7% (SEQ ID NO: 226)150 mL/Ha 16.7 μM 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BCpreservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition13) Bt.4Q7Flg22 4.11 Fl. oz/Ac or +427.2 kg/Ha 136.1% (SEQ ID NO: 226)300 mL/Ha 16.7 μM 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BCpreservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition14) Gm.RHPP 2.05 Fl. oz/Ac or +294.2 kg/Ha 125.5% (SEQ ID NO: 600) 150mL/Ha 100 μM PROXEL BC preservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM(MIT) (Composition 15) Gm.RHPP 4.11 Fl. oz/Ac or +506.8 kg/Ha 143.5%(SEQ ID NO: 600) 300 mL/Ha 100 μM PROXEL BC preservative: 330.7 μM; 50.1μM (CMIT); 21.71 μM (MIT) (Composition 16) FOX Fungicide + 5.48 Fl.oz/Ac or +426.5 kg/Ha 138.4% Bt.4Q7Flg22 400 mL/Ha + (SEQ ID NO: 226)2.05 Fl. oz/Ac or 16.7 μM 150 mL/Ha 1.67 mM Sodium Phosphate Buffer, pH5.7 PROXEL BC preservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM(MIT) (Composition 17) FOX Fungicide + 5.48 Fl. oz/Ac or +418.5 kg/Ha137.2% Bt.4Q7Flg22 400 mL/Ha + (SEQ ID NO: 226) 4.11 Fl. oz/Ac or 16.7μM 300 mL/Ha 1.67 mM Sodium Phosphate Buffer, pH 5.7 PROXEL BCpreservative: 330.7 μM (BIT); 53.5 μM (CMIT); 26.1 μM (MIT) (Composition18) FOX Fungicide + 5.48 Fl. oz/Ac or +517.6 kg/Ha 145.5% Gm.RHPP 400mL/Ha + (SEQ ID NO: 600) 2.05 Fl. oz/Ac or 100 μM 150 mL/Ha PROXEL BCpreservative: 330.7 μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 19)FOX Fungicide + 5.48 Fl. oz/Ac or +539.9 kg/Ha 146.9% Gm.RHPP 400mL/Ha + (SEQ ID NO: 600) 300 mL/Ha 100 μM PROXEL BC preservative: 330.7μM; 50.1 μM (CMIT); 21.71 μM (MIT) (Composition 20)

Example 57. Treatment of Citrus Trees Infected with Candidatusliberibacter asiaticus with Flg22 Increases Fruit Set

Previous results summarized in Example 51 indicate that Bt.4Q7Flg22 (SEQID NO: 226) trunk injection reduces pathogen titer and promotes newgrowth in citrus trees infected with Candidatus liberibacter asiaticus,the causative agent of Huanglongbing (HLB). To assess for a potentialincrease in fruiting and obtain early estimates of yield, fruit set wasmeasured in June 2018 for the same HLB-infected ‘Valencia’ Orange (to beharvested spring 2019) and ‘Ruby Red’ Grapefruit trees (to be harvestedfall 2018) that were trunk-injected with Bt.4Q7Flg22 in April 2017 atthe commercial grove orchard located in central Florida (Okeechobeecounty). As described in Example 51, trees were injected in April 2017with either a 1× Bt.4Q7Flg22-Low Rate (0.55 micromoles peptide; 0.138 μMestimated phloem concentration) or a 10× Bt.4Q7Flg22-High Rate (5.5micromoles peptide; 1.38 μM estimated phloem concentration). In June2018, the Bt.4Q7Flg22-injected trees were compared to untreated controltrees within the same area of the grove using established methods forprojecting citrus tree yield (“Forecasting Florida Citrus Production:Methodology & Development; 1971; by S. R. Williams for Florida Crop andLivestock Reporting Service). To quantify fruit set, three quaternarylimbs at eye level were randomly chosen on each tree (n=8 trees pertreatment ‘Valencia’ orange, n=10 trees per treatment ‘Ruby Red’grapefruit). The circumference of each quaternary limb was measured atthe junction where the limb began and used to calculate thecross-sectional area (CSA) of the limb using the following equations(where C=circumference, CSA=cross-sectional area, and r=radius):

$r = {{\frac{c}{2\; \pi}\mspace{14mu} {and}\mspace{14mu} {CSA}} = {\pi \; r^{2}}}$

Then, the total number of fruit on the quaternary limb distal to thatjunction were counted. To normalize for limb size, fruit set for eachquaternary limb was quantified as the number of fruit on the limbdivided by the CSA of the quaternary limb:

${{Fruit}\mspace{14mu} {set}} = \frac{{Total}\mspace{14mu} {fruit}\mspace{14mu} {per}\mspace{14mu} {limb}}{{Limb}\mspace{14mu} {CSA}}$

The fruit count per quaternary limb CSA is reported in FIG. 11(‘Valencia’ orange) and FIG. 12 (Red Grapefruit) in box and whiskerplots, where the median value for each treatment is marked as thevertical line within the box, the mean or average value is marked by the“x”, the upper and lower quartiles are marked by the ends of the box,and the whiskers extend to the highest and lowest observed fruit countsper limb CSA. Any outlier values are indicated by the small circleslocated outside the standard error bars for each treatment.

To further assess the size and volume of fruit setting per tree, thefruit diameter (mm) of at least 10 randomly chosen fruit per tree wasmeasured using calipers placed at the widest point on each fruit. Theaverage fruit diameter (mm) per tree for each treatment is reported inFIG. 13 (‘Valencia’ orange) and FIG. 14 (Red Grapefruit) in box andwhisker plots. The average fruit diameter was used to estimate the totalfruit volume per limb for each treatment. For these estimates, thevolume in milliliters (mL) of a theoretically spherical orange wascalculated using the following equation, where the radius (r) of thefruit is the average diameter (measured in mm) per limb divided by 2:

${{Total}\mspace{14mu} {F{ruit}}\mspace{14mu} {Volume}\mspace{14mu} {per}\mspace{14mu} {limb}\mspace{14mu} ({mL})} = {{Total}\mspace{14mu} {fruit}\mspace{14mu} {per}\mspace{14mu} {limb}*\frac{4}{3}\pi \; r^{3}*\frac{1\mspace{14mu} {mL}}{1000\mspace{14mu} {cubic}\mspace{14mu} {millimeters}}}$

The estimated volume of fruit normalized by limb CSA for each treatmentis reported in FIG. 15 (‘Valencia’ orange) and FIG. 16 (Red Grapefruit)in box and whisker plots.

The measurements collected in June 2018 to assess fruit set in‘Valencia’ orange and ‘Ruby Red’ grapefruit trees in Okeechobee, Fla.show increased fruit per limb and increased fruit size for trees of bothvarieties receiving trunk injections of 1× Low and 10× High rates ofBt.4Q7Flg22 (SEQ ID NO: 226) in April 2017, when comparing the mean andmedian values for all parameters measured versus the untreated control.The increased fruit set and size are predicative of increased yield.These results provide further evidence that trunk injection of citrustrees with Bt.4Q7Flg22 can be utilized to reduce C. liberibacterbacterial titers in orange (FIG. 9) and grapefruit (FIG. 10; Table 84)and stimulate new shoot and fruit growth (Table 85, FIGS. 11-16) incitrus trees.

In view of the above, it will be seen that the several objects of theinvention are achieved and other advantageous results attained.

As various changes could be made in the above polypeptides, recombinantorganisms, methods, and seeds, without departing from the scope of theinvention, it is intended that all matter contained in the abovedescription shall be interpreted as illustrative and not in a limitingsense.

EMBODIMENTS

For further illustration, additional non-limiting embodiments of thepresent disclosure are set forth below.

Embodiment 1 is a polypeptide for bioactive priming of a plant or aplant part to increase growth, yield, health, longevity, productivity,and/or vigor of a plant or a plant part and/or decrease abiotic stressin the plant or the plant part and/or protect the plant or the plantpart from disease, insects and/or nematodes, and/or increase the innateimmune response of the plant or the plant part and/or change plantarchitecture, wherein the polypeptide comprises:

(a) a flagellin or flagellin-associated polypeptide and an amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 1-225, 227-375, 526, 528, 530, 532, 534,536, 538, 540, 541, 751, 752, and 754-766; or

(b) a mutant flagellin or flagellin-associated polypeptide and an aminoacid sequence of the mutant flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 571-579, and 753; or

(c) a mutant flagellin or flagellin-associated polypeptide and an aminoacid sequence of the mutant flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 580-586; or

(d) a retro inverso Flg22 polypeptide and an amino acid sequence of theretro inverso Flg22 polypeptide comprises any one of SEQ ID NOs:376-450, 527, 531, 533, 535, 537 and 539; or

(e) a retro inverso FlgII-28 polypeptide and an amino acid sequence ofthe retro inverso FlgII-28 polypeptide comprises any one of SEQ ID NOs:451-525, or 768; or

(f) a retro inverso Flg15 polypeptide and an amino acid sequence of theretro inverso Flg15 polypeptide comprises SEQ ID NOs: 529 or 767; or

(g) a harpin or harpin-like polypeptide and an amino acid sequence ofthe harpin or harpin-like polypeptide comprises any one of SEQ ID NOs:587, 589, 591, 593, 594 and 595; or

(h) a retro inverso harpin or harpin-like polypeptide and an amino acidsequence of the retro inverso harpin or harpin-like polypeptidecomprises any one of SEQ ID NOs: 588, 590, 592, 596 and 597; or

(i) a root hair promoting polypeptide (RHPP) and an amino acid sequenceof the RHPP comprises any one of SEQ ID Nos: 600, 603 and 604; or

(j) a Kunitz Trypsin Inhibitor (KTI) polypeptide and an amino acidsequence of the KTI polypeptide comprises SEQ ID No: 602; or

(k) a retro inverso root hair promoting polypeptide (RI RHPP) and anamino acid sequence of the RI RHPP comprises any one of SEQ ID NO: 601,605 and 606; or

(l) an elongation factor Tu (EF-Tu) polypeptide and an amino acidsequence of the EF-Tu polypeptide comprises any one of SEQ ID NOs:607-623; or

(m) a retro inverso elongation factor Tu (RI EF-Tu) polypeptide and anamino acid sequence of the RI EF-Tu polypeptide comprises any one of SEQID NOs: 624-640; or

(n) a fusion polypeptide comprising SEQ ID NO: 750; or

(o) a phytosulfokine (PSK) polypeptide and an amino acid sequence of thePSK polypeptide comprises SEQ ID NO: 598; or

(p) a retro inverso phytosulfokine (RI PSK) polypeptide and an aminoacid sequence of the RI PSK polypeptide comprises SEQ ID NO: 599; or

(q) a thionin or thionin-like polypeptide and an amino acid sequence ofthe thionin or thionin-like polypeptide comprises any one of SEQ ID NOs:650-749, and optionally, wherein the flagellin or flagellin-associatedpolypeptide of (a), the mutant flagellin or flagellin-associatedpolypeptide of (c), the harpin or harpin-like polypeptide of (g), theKTI polypeptide of (j), the PSK polypeptide of (o), and the thionin orthionin-like polypeptide of (q) either: contains a chemicalmodification; is a variant having an amino acid insertion, deletion,inversion, repeat, duplication, extension, or substitution within theamino acid; is part of a fusion protein; or contains a proteaserecognition sequence.

Embodiment 2 is a polypeptide of embodiment 1, wherein the flagellin orflagellin-associated polypeptide of (a), the mutant flagellin orflagellin-associated polypeptide of (c), the harpin or harpin-likepolypeptide of (g), the KTI polypeptide of (j), the PSK polypeptide of(o), and the thionin or thionin-like polypeptide of (q) either: containsa chemical modification; is a variant having an amino acid insertion,deletion, inversion, repeat, duplication, extension, or substitutionwithin the amino acid; is part of a fusion protein; or contains aprotease recognition sequence.

Embodiment 3 is a polypeptide of embodiment 2, wherein the chemicalmodification comprises acetylation, acid addition, acylation,ADP-ribosylation, aldehyde addition, alkylamide addition, amidation, amination, biotinylation, carbamate addition, chloromethyl ketoneaddition, covalent attachment of a nucleotide or nucleotide derivative,cross-linking, cyclization, disulfide bond formation, demethylation,ester addition, formation of covalent cross-links, formation ofcysteine-cysteine disulfide bonds, formation of pyroglutamate,formylation, gamma-carboxylation, glycosylation, GPI anchor formation,hydrazide addition, hydroxyamic acid addition, hydroxylation,iodination, lipid addition, methylation, myristoylation, oxidation,PEGylation, proteolytic processing, phosphorylation, prenylation,palmitoylation, addition of a purification tag, pyroglutamyl addition,racemization, selenoylation, sulfonamide addition, sulfation,transfer-RNA mediated addition of amino acids to proteins,ubiquitination, or urea addition.

Embodiment 4 is a polypeptide of embodiment 2, wherein the chemicalmodification comprises an N-terminal modification or a C-terminalmodification.

Embodiment 5 is a polypeptide of embodiment 3, wherein the chemicalmodification comprises acetylation, amidation, cross-linking, orcyclization.

Embodiment 6 is a polypeptide of embodiment 2, wherein the amino acidsubstitution within the amino acid of the variant comprises substitutionof a β-amino acid, a D-amino acid, or a non-natural amino acid.

Embodiment 7 is a polypeptide of embodiment 1, wherein the polypeptidecomprises: the mutant flagellin or flagellin-associated polypeptide of(b); or the retro inverso Flg22 polypeptide of (d); or the retro inversoFlgII-28 polypeptide of (e); or the retro inverso Flg15 polypeptide of(f); or the retro inverso harpin or harpin-like polypeptide of (h); orthe RHPP of (i); or the RI RHPP of (k); or the EF-Tu polypeptide of (l);or the RI EF-Tu polypeptide of (m); or the fusion polypeptide of (n); orthe RI PSK polypeptide of (p).

Embodiment 8 is a polypeptide of embodiment 7, wherein the amino acidsequence of the mutant flagellin or flagellin-associated polypeptide of(b) comprises any one of SEQ ID NOs: 571-573.

Embodiment 9 is a polypeptide of any one of embodiments 1-6, wherein thepolypeptide comprises the flagellin or flagellin-associated polypeptide.

Embodiment 10 is a polypeptide of embodiment 9, wherein the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs 754-766.

Embodiment 11 is a polypeptide of any one of embodiments 1-6, whereinthe polypeptide comprises the mutant flagellin or flagellin-associatedpolypeptide.

Embodiment 12 is a polypeptide of embodiment 11, wherein the mutantflagellin or flagellin-associated polypeptide is from a Bacillus, aLysinibacillus, a Paenibacillus, or an Aneurinibacillus genus bacterium.

Embodiment 13 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the retro inverso Flg22 polypeptide.

Embodiment 14 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the retro inverso FlgII-28 polypeptide.

Embodiment 15 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the retro inverso Flg15 polypeptide.

Embodiment 16 is a polypeptide of any one of embodiments 1-6, whereinthe polypeptide comprises the harpin or harpin-like polypeptide.

Embodiment 17 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the retro inverso harpin or harpin-like polypeptide.

Embodiment 18 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the RHPP.

Embodiment 19 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the RI RHPP.

Embodiment 20 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the EF-Tu polypeptide.

Embodiment 21 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the retro inverso EF-Tu polypeptide.

Embodiment 22 is a polypeptide of any one of embodiments 1-21, whereinthe polypeptide further comprises a core sequence.

Embodiment 23 is a polypeptide of embodiment 22 wherein the coresequence comprises any one of SEQ IDs 754-766.

Embodiment 24 is a polypeptide of embodiment 1, wherein the polypeptidecomprises the fusion polypeptide.

Embodiment 25 is a polypeptide of embodiment 24, wherein the fusionpolypeptide comprises an assistance polypeptide.

Embodiment 26 is a polypeptide of embodiment 25, wherein the assistancepolypeptide comprises a signature polypeptide, and an amino acidsequence of the signature polypeptide comprises any one of SEQ ID NOs:542-548, or any combination thereof.

Embodiment 27 is a polypeptide of embodiment 25, wherein the assistancepolypeptide comprises a signature polypeptide, and an amino acidsequence of the signature polypeptide comprises SEQ ID NO: 542.

Embodiment 28 is a polypeptide of embodiment 25, wherein the assistancepolypeptide comprises a signal anchor sorting polypeptide, and an aminoacid sequence of the signal anchor sorting polypeptide comprises any oneof SEQ ID NOs: 549-562, or any combination thereof.

Embodiment 29 is a polypeptide of embodiment 25, wherein the assistancepolypeptide comprises a signal anchor sorting polypeptide, and an aminoacid sequence of the signal anchor sorting polypeptide comprises SEQ IDNO: 549.

Embodiment 30 is a polypeptide of any one of embodiments 1-6, whereinthe polypeptide comprises the phytosulfokine polypeptide.

Embodiment 31 is a polypeptide of any one of embodiments 1-6, whereinthe polypeptide comprises the retro inverso phytosulfokine polypeptide.

Embodiment 32 is a polypeptide of any one of embodiments 1-6, whereinthe polypeptide comprises the thionin or thionin-like polypeptide.

Embodiment 33 is a polypeptide of any one of embodiment 1-32, whereinthe polypeptide further comprises a core sequence comprising any one ofSEQ ID NOs: 754-768, and wherein the inclusion of the core sequenceincreases the bioactive priming activity of the polypeptide.

Embodiment 34 is a polypeptide of any one of embodiments 1-33, whereinthe polypeptide can comprise an amino acid sequence having at least 70%identity to any one of SEQ ID NOs. 1-768, and the polypeptide hasbioactive priming activity.

Embodiment 35 is a polypeptide of any one of embodiments 1-34, whereinthe polypeptide can comprise an amino acid sequence having at least 75%identity to any one of SEQ ID NOs. 1-768, and the polypeptide hasbioactive priming activity.

Embodiment 36 is a polypeptide of any one of embodiments 1-35, whereinthe polypeptide can comprise an amino acid sequence having at least 80%identity to any one of SEQ ID NOs. 1-768, and the polypeptide hasbioactive priming activity.

Embodiment 37 is a polypeptide of any one of embodiments 1-36, whereinthe polypeptide can comprise an amino acid sequence having at least 85%identity to any one of SEQ ID NOs. 1-768, and the polypeptide hasbioactive priming activity.

Embodiment 38 is a polypeptide of any one of embodiments 1-37, whereinthe polypeptide can comprise an amino acid sequence having at least 90%identity to any one of SEQ ID NOs. 1-768, and the polypeptide hasbioactive priming activity.

Embodiment 39 is a polypeptide of any one of embodiments 1-38, whereinthe polypeptide can comprise an amino acid sequence having at least 95%identity to any one of SEQ ID NOs. 1-768, and the polypeptide hasbioactive priming activity.

Embodiment 40 is a polypeptide of any one of embodiments 1-39, whereinthe polypeptide can comprise an amino acid sequence having at least 98%identity to any one of SEQ ID NOs. 1-768, and the polypeptide hasbioactive priming activity.

Embodiment 41 is a polypeptide of any one of embodiments 1-40, whereinthe polypeptide can comprise an amino acid sequence having at least 99%identity to any one of SEQ ID NOs. 1-766, and the polypeptide hasbioactive priming activity.

Embodiment 42 is a polypeptide of any one of embodiments 1-41, whereinthe polypeptide is isolated, concentrated from a fermentation product,and/or partially purified.

Embodiment 43 is a polypeptide of embodiment 42 wherein the polypeptideis isolated, concentrated from a fermentation product, and/or partiallypurified by filtration or chromatography.

Embodiment 44 is a polypeptide of embodiment 42 or 43 wherein thepolypeptide is isolated, concentrated from a fermentation product, orpartially purified from a recombinant microorganism.

Embodiment 45 is a composition for bioactive priming of a plant or aplant part to increase growth, yield, health, longevity, productivity,and/or vigor of a plant or a plant part and/or decrease abiotic stressin the plant or the plant part and/or protect the plant or the plantpart from disease, insects and/or nematodes, and/or increase the innateimmune response of the plant or the plant part and/or change plantarchitecture, the composition comprising either:

the polypeptide of any one of embodiments 1-44, or any combinationthereof, and an agrochemical or a carrier; or

any combination of the polypeptides of any one of embodiments 1-44.

Embodiment 46 is a composition of embodiment 45, wherein the compositioncomprises the flagellin or flagellin-associated polypeptide of (a), themutant flagellin or flagellin-associated polypeptide of (c), the harpinor harpin-like polypeptide of (g), the KTI polypeptide of (j), the PSKpolypeptide of (o), the thionin or thionin-like polypeptide of (q), orany combination thereof, and an agrochemical or carrier, theagrochemical or carrier not being associated with the polypeptide innature.

Embodiment 47 is a composition of embodiment 45, wherein the compositioncomprises:

the mutant flagellin or flagellin-associated polypeptide of (b); or theretro inverso Flg22 polypeptide of (d); or the retro inverso FlgII-28polypeptide of (e); or the retro inverso Flg15 polypeptide of (f); orthe retro inverso harpin or harpin-like polypeptide of (h); or the RHPPof (i); or the RI RHPP of (k); or the EF-Tu polypeptide of (l); or theRI EF-Tu polypeptide of (m); or the fusion polypeptide of (n); the RIPSK polypeptide of (p), or any combination thereof; and

the flagellin or flagellin-associated polypeptide of (a), the mutantflagellin or flagellin-associated polypeptide of (c), the harpin orharpin-like polypeptide of (g), the KTI polypeptide of (j), the PSKpolypeptide of (o), the thionin or thionin-like polypeptide of (q), orany combination thereof.

Embodiment 48 is a composition of embodiment 46 or 47, wherein theflagellin or flagellin-associated polypeptide of (a), the mutantflagellin or flagellin-associated polypeptide of (c), the harpin orharpin-like polypeptide of (g), the KTI polypeptide of (j), the PSKpolypeptide of (o), the thionin or thionin-like polypeptide of (q), orany combination thereof does not contain the chemical modification; isnot the variant; is not part of the fusion protein; and does not containthe protease recognition sequence.

Embodiment 49 is a composition of embodiment 45, wherein the compositioncomprises: two or more of the mutant flagellin or flagellin-associatedpolypeptide of (b); or the retro inverso Flg22 polypeptide of (d); orthe retro inverso FlgII-28 polypeptide of (e); or the retro inversoFlg15 polypeptide of (f); or the retro inverso harpin or harpin-likepolypeptide of (h); or the RHPP of (i); or the RI RHPP of (k); or theEF-Tu polypeptide of (l); or the RI EF-Tu polypeptide of (m); or thefusion polypeptide of (n); or the RI PSK polypeptide of (p).

Embodiment 50 is a composition of embodiment 45, wherein the compositioncomprises: two or more of the flagellin or flagellin-associatedpolypeptide of (a), the mutant flagellin or flagellin-associatedpolypeptide of (c), the harpin or harpin-like polypeptide of (g), theKTI polypeptide of (j), the PSK polypeptide of (o), or the thionin orthionin-like polypeptide of (q), wherein either:

at least one of the flagellin or flagellin-associated polypeptide of(a), the mutant flagellin or flagellin-associated polypeptide of (c),the harpin or harpin-like polypeptide of (g), the PSK polypeptide of(o), or the thionin or thionin-like polypeptide of (q) contains thechemical modification; is the variant; is part of the fusion protein; orcontains the protease recognition sequence; or

at least two of the flagellin or flagellin-associated polypeptide of(a), the mutant flagellin or flagellin-associated polypeptide of (c),the harpin or harpin-like polypeptide of (g), the KTI polypeptide of(j), the PSK polypeptide of (o), or the thionin or thionin-likepolypeptide of (q) are not associated in nature.

Embodiment 51 is a composition of embodiment 45, wherein the flagellinor flagellin-associated polypeptide is produced recombinantly by amicroorganism.

Embodiment 52 is a composition of embodiment 51, wherein themicroorganism comprises a bacterium from the genus Bacillus, a bacteriumfrom the genus Pseudomonas, a bacterium from the genus Paenibacillus, afungus of the genus Penicillium, a bacterium of the genus Glomus, abacterium of the genus Arthrobacter, a bacterium of the genusParacoccus, a bacterium of the genus Rhizobium, a bacterium of the genusBradyrhizobium, a bacterium of the genus Azosprillium, a bacterium ofthe genus Enterobacter, a bacterium of the genus Escherichia, or anycombination thereof.

Embodiment 53 is a recombinant microorganism that expresses oroverexpresses a polypeptide, wherein the polypeptide comprises thepolypeptide of any one of embodiments 1-44 or any combination thereof.

Embodiment 54 is a recombinant microorganism of embodiment 53, whereinthe polypeptide comprises: the flagellin or flagellin-associatedpolypeptide of (a); or the mutant flagellin or flagellin-associatedpolypeptide of (b); or the mutant flagellin or flagellin-associatedpolypeptide of (c); or the harpin or harpin-like polypeptide of (g); orthe RHPP of (i); or the KTI polypeptide of (j); or the EF-Tu polypeptideof (l); or the fusion polypeptide of (n); or the PSK polypeptide of (o);or the thionin or thionin-like polypeptide of (q).

Embodiment 55 is a recombinant microorganism of embodiment 53 or 54,wherein the polypeptide is overexpressed by the microorganism.

Embodiment 56 is a recombinant microorganism of embodiment 55, whereinthe polypeptide is made with a secretion signal.

Embodiment 57 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the flagellin orflagellin-associated polypeptide.

Embodiment 58 is a composition or recombinant microorganism ofembodiment 57, wherein the flagellin or flagellin-associated polypeptideis modified chemically on its N or C terminus.

Embodiment 59 is a composition or recombinant microorganism ofembodiment 57, wherein the flagellin or flagellin-associated polypeptideis modified via crosslinking or cyclization.

Embodiment 60 is a composition or recombinant microorganism ofembodiment 57, wherein the flagellin or flagellin-associated polypeptideis from a Bacillus, a Lysinibacillus, a Paenibacillus, anAneurinibacillus genus bacterium, or any combination thereof.

Embodiment 61 is a composition or recombinant microorganism ofembodiment 57, wherein the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs: 1-75,or any combination thereof.

Embodiment 62 is a composition or recombinant microorganism ofembodiment 57, wherein the flagellin or flagellin-associated polypeptidecomprises a truncated N-terminal polypeptide and an amino acid sequenceof the truncated N-terminal polypeptide comprises SEQ ID NO: 76, 78, 80,82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 109, 110,112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138,140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166,168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194,196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222,224, 752, or any combination thereof.

Embodiment 63 is a composition or recombinant microorganism ofembodiment 57, wherein the flagellin or flagellin-associated polypeptidecomprises a truncated C-terminal polypeptide and an amino acid sequenceof the truncated C-terminal polypeptide comprises SEQ ID NO: 77, 79, 81,83, 85, 87, 89, 91, 93, 95, 97, 99, 101, 103, 105, 107, 109, 111, 113,115, 117, 119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 141, 143,145, 147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171,173, 175, 177, 179, 181, 183, 185, 187, 189, 191, 201, 203, 205, 207,209, 211, 213, 215, 217, 219, 221, 223, 225, or any combination thereof.

Embodiment 64 is a composition or recombinant microorganism ofembodiment 57, wherein the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs: 226,289, 290, 291, 293, 294, 295, 300, 437, 532, 534, 536, 538, 540,571-586, and 751-768 or any combination thereof.

Embodiment 65 is a composition or recombinant microorganism ofembodiment 64, wherein the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs: 226,293, 295, 300, 540, 571, 574 and 752, or any combination thereof.

Embodiment 66 is a composition or recombinant microorganism ofembodiment 64 wherein the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs754-766.

Embodiment 67 is a composition or recombinant microorganism ofembodiment 66 wherein the flagellin or flagellin-associated polypeptideis incorporated into a non-flagellin or flagellin-associated polypeptideor full length protein, wherein the incorporation of the flagellin orflagellin-associated polypeptide results in increased bioactive primingactivity in the non-flagellin or flagellin-associated polypeptide orfull length protein.

Embodiment 68 is a composition or recombinant microorganism ofembodiment 57, wherein the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs:226-300, or any combination thereof.

Embodiment 69 is a composition or recombinant microorganism ofembodiment 57, wherein the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs:301-375, and 751 or any combination thereof.

Embodiment 70 is a composition or recombinant microorganism ofembodiment 69, wherein the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises SEQ ID NO: 301.

Embodiment 71 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the retroinverso Flg22 polypeptide.

Embodiment 72 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the retroinverso FlgII-28 polypeptide.

Embodiment 73 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the thionin orthionin-like polypeptide.

Embodiment 74 is a composition or recombinant microorganism ofembodiment 73, wherein the amino acid sequence of the thionin orthionin-like polypeptide comprises SEQ ID NO: 651.

Embodiment 75 is a composition or recombinant microorganism ofembodiment 74, wherein the thionin or thionin-like polypeptide is fusedto a phloem targeting sequence to form a fused polypeptide, the aminoacid sequence of the phloem targeting sequence comprising any one of SEQID NOs: 641-649, or any combination thereof, for delivering the fusedpolypeptide to vascular tissue or cells and/or phloem orphloem-associated tissue or cells in the plant or plant part.

Embodiment 76 is a composition or recombinant microorganism ofembodiment 75, wherein the amino acid sequence of the phloem targetingsequence comprises SEQ ID NO: 641.

Embodiment 77 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the PSKpolypeptide.

Embodiment 78 is a composition or recombinant microorganism ofembodiment 77, wherein the amino acid sequence of the PSK polypeptidecomprises SEQ ID NO: 598.

Embodiment 79 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the RHPP.

Embodiment 80 is a composition or recombinant microorganism ofembodiment 79, wherein the amino acid sequence of the RHPP comprises SEQID NO: 600.

Embodiment 81 is a composition or recombinant microorganism ofembodiment 80, further comprising the flagellin or flagellin-associatepolypeptide.

Embodiment 82 is a composition or recombinant microorganism ofembodiment 81, wherein the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs: 226,752, and 571.

Embodiment 83 is a composition or recombinant microorganism ofembodiment 82, wherein the amino acid sequence of the flagellin orflagellin associated polypeptide comprises SEQ ID NO: 226.

Embodiment 84 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the RI RHPP.

Embodiment 85 is a composition or recombinant microorganism of any oneof embodiments 38-47, wherein the polypeptide comprises the harpin orharpin-like polypeptide.

Embodiment 86 is a composition or recombinant microorganism ofembodiment 85, wherein the amino acid sequence of the harpin orharpin-like polypeptide comprises SEQ ID NO: 587.

Embodiment 87 is a composition or recombinant microorganism of any oneof embodiments 45-56, comprising the PSK polypeptide, the RHPP, theharpin or harpin-like polypeptide, or a combination thereof.

Embodiment 88 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the EF-Tupolypeptide.

Embodiment 89 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide further comprises asecretion signal.

Embodiment 90 is a composition or recombinant microorganism ofembodiment 88 or 89, wherein the amino acid sequence of the EF-Tupolypeptide comprises SEQ ID NO: 616.

Embodiment 91 is a composition or recombinant microorganism ofembodiment 90, further comprising an EF-Tu polypeptide having an aminoacid sequence comprising SEQ ID NO: 617.

Embodiment 92 is a composition or recombinant microorganism of any oneof embodiments 88-91, further comprising the flagellin orflagellin-associated polypeptide, the amino acid sequence of theflagellin or flagellin-associated polypeptide comprising any one of SEQID NOs: 1-525, 526, 530, 532, 534, 536, 538, 540, 541, 571-586, and751-766, or any combination thereof.

Embodiment 93 is a composition or recombinant microorganism ofembodiment 92, further comprising the flagellin or flagellin-associatedpolypeptide, the amino acid sequence of the flagellin orflagellin-associated polypeptide comprising SEQ ID NO: 226, 571 or 752or a combination thereof.

Embodiment 94 is a composition or recombinant microorganism ofembodiment 92, further comprising the harpin or harpin-like polypeptide.

Embodiment 95 is a composition or recombinant microorganism ofembodiment 94, wherein amino acid sequence of the harpin or harpin-likepolypeptide comprises SEQ ID NO: 587.

Embodiment 96 is a composition or recombinant microorganism of any oneof embodiments 45-56, comprising

(a) the flagellin or flagellin-associated polypeptides and the aminoacid sequences of the flagellin or flagellin-associated polypeptidescomprise SEQ ID NOs: 571, 295, 300, 293, and 580; or 295, 300, 293, and580; or 571, 295, 293, and 580; or 571, 300, 293, and 580; or 571, 293and 580; or 571, 295, 293; or

(b) the flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226 and one or more of cellobiose, chitin, chitosan, orcellulose; or

(c) the flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226 and the harpin or harpin-like polypeptide and the aminoacid sequence of the harpin or harpin-like polypeptide comprises SEQ IDNO: 591; or

(d) the harpin or harpin-like polypeptide and the amino acid sequence ofthe harpin or harpin-like polypeptide comprises SEQ ID NO: 587 and thePSK polypeptide and the amino acid sequence of the PSK polypeptidecomprises SEQ ID NO: 598; or

(e) the flagellin or flagellin-associated polypeptide and the amino acidsequence—of the flagellin or flagellin-associated polypeptide—comprisesSEQ ID NO: 226, 571 or 752 or any combination thereof and the EF-Tupolypeptides and the amino acid sequences of the EF-Tu polypeptidescomprise SEQ ID NOs: 616 and 617; or

(f) the flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226, 540, 571 or 752, or any combination thereof; or

(g) the RHPP polypeptide and the amino acid sequence of the RHPPpolypeptide comprises SEQ ID NO: 600; or

(h) the flagellin or flagellin-associated polypeptide and the amino acidsequences of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226, 540, 226, 571, or 752, or any combination thereof andthe RHPP polypeptide and the amino acid sequence of the RHPP polypeptidecomprises SEQ ID NO: 600; or

(i) the flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226 and the RHPP polypeptide and the amino acid sequence ofthe RHPP polypeptide comprises SEQ ID NO: 600.

Embodiment 97 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the polypeptide comprises the fusionpolypeptide.

Embodiment 98 is a composition or recombinant microorganism of any oneof embodiments 45-56, wherein the flagellin or flagellin-associatedpolypeptide is a fusion protein with a polypeptide.

Embodiment 99 is a composition or recombinant microorganism of any oneof embodiments 45-98, further comprising an assistance polypeptide.

Embodiment 100 is a composition or recombinant microorganism ofembodiment 99, wherein the assistance polypeptide comprises a signaturepolypeptide, and an amino acid sequence of the signature polypeptidecomprises any one of SEQ ID NOs: 542-548, or any combination thereof.

Embodiment 101 is a composition or recombinant microorganism ofembodiment 99, wherein the assistance polypeptide comprises a signaturepolypeptide, and an amino acid sequence of the signature polypeptidecomprises SEQ ID NO: 542.

Embodiment 102 is a composition or recombinant microorganism ofembodiment 99, wherein the assistance polypeptide comprises a signalanchor sorting polypeptide, and an amino acid sequence of the signalanchor sorting polypeptide comprises any one of SEQ ID NOs: 549-562, orany combination thereof.

Embodiment 103 is a composition or recombinant microorganism ofembodiment 102, wherein the assistance polypeptide comprises a signalanchor sorting polypeptide, and an amino acid sequence of the signalanchor sorting polypeptide comprises SEQ ID NO: 549.

Embodiment 104 is a composition or recombinant microorganism ofembodiment 99, wherein the assistance polypeptide comprises a secretionpolypeptide, and an amino acid sequence of the secretion polypeptidecomprises any one of SEQ ID NOs: 563-570, or 769, or any combinationthereof.

Embodiment 105 is a composition or recombinant microorganism ofembodiment 104, wherein the assistance polypeptide comprises a secretionpolypeptide, and an amino acid sequence of the secretion polypeptidecomprises SEQ ID NO: 563 or 769.

Embodiment 106 is a composition or recombinant microorganism ofembodiment 99 or 104, wherein the assistance polypeptide comprises anenterokinase cleavage sequence and an amino acid sequence of theenterokinase cleavage sequence comprises SEQ ID NO: 772.

Embodiment 107 is a composition or recombinant microorganism ofembodiment 106, wherein assistance polypeptide comprises a linkingregion between the polypeptide and a protein tag.

Embodiment 108 is a composition or recombinant microorganism ofembodiment 107, wherein the protein tag comprises a poly-histidine (His)tag, a FLAG tag, an antibody epitope, streptavidin/biotin, glutathioneS-transferase (GST), or any combination thereof.

Embodiment 109 is a composition of any one of embodiments 45-52 and57-108, further comprising either an agrochemical or a carrier which isassociated with the polypeptide in nature.

Embodiment 110 is a composition of any one of embodiments 45-52 and57-109, wherein the agrochemical comprises an antibiotic, abiopesticide, a preservative, a buffering agent, a wetting agent, asurfactant, a coating agent, a monosaccharide, a polysaccharide, anabrading agent, a pesticide, an insecticide, a herbicide, a nematicide,a bacteriocide, a fungicide, a miticide, a fertilizer, a biostimulant,an osmoprotectant, a colorant, a humectant, an amino acid, a biologicalcontrol agent, or a combination thereof.

Embodiment 111 is a composition of embodiment 110, wherein thepreservative comprises dichlorophene, benzylalcohol hemi formal, anisothiazolinone derivative, an alkylisothiazolinone, abenzisothiazolinone, MIT (2-methyl-4-isothiazolin-3-one), BIT(1,2-benzisothiazolin-3-one),5-chloro-2-(4-chlorobenzyl)-3(2H)-isothiazolone,5-chloro-2-methyl-2H-isothiazol-3-one,5-chloro-2-methyl-2H-isothiazol-3-one,5-chloro-2-methyl-2H-isothiazol-3-one-hydrochloride,4,5-dichloro-2-cyclohexyl-4-isothiazolin-3-one,4,5-dichloro-2-octyl-2H-isothiazol-3-one, 2-methyl-2H-isothiazol-3-one,2-methyl-2H-isothiazol-3-one-calcium chloride complex,2-octyl-2H-isothiazol-3-one, or any combination thereof.

Embodiment 112 is a composition of embodiment 110 or 111, wherein thebuffering agent comprises potassium, phosphoric acid, a phosphate salt,citric acid, a citrate salt, a sulfate salt, MOPS, HEPES, or anycombination thereof.

Embodiment 113 is a composition of any one of embodiments 110-112,wherein the wetting agent comprises an organosilicone, apolyoxyethoxylate, a polysorbate, a polyethyleneglycol or derivativethereof, an ethoxylate, a crop oil, a polysaccharide, or any combinationthereof.

Embodiment 114 is a composition of any one of embodiments 110-113,wherein the surfactant comprises a heavy petroleum oil, a heavypetroleum distillate, a polyol fatty acid ester, a polyethoxylated fattyacid ester, an aryl alkyl polyoxyethylene glycol, an alkyl amineacetate, an alkyl aryl sulfonate, a polyhydric alcohol, an alkylphosphate, an alcohol ethoxylate, an alkylphenol ethoxylate, alkoxylatedpolyols, alkyl polyethoxy ethers, ethoxylated soybean oil derivatives,alkylpolyoxyehylene glycerol, alcohol ethoxylate,polyoxyehylenepolyoxypropylene monobutyl ether, an organosiliconederivative, or any combination thereof.

Embodiment 115 is a composition of any one of embodiments 110-114,wherein the coating agent comprises a tackifier, a polymers, a fillingagent, a bulking agent, or any combination thereof.

Embodiment 116 is a composition of embodiment 115, wherein the tackifiercomprises carboxymethylcellulose, a natural polymer, a syntheticpolymer, gum Arabic, chitin, polyvinyl alcohol, polyvinyl acetate, anatural phospholipid, a cephalin, a lecithin, a synthetic phospholipid,a polyester, a polyether ester, a polyanhydride, a polyester urethane, apolyester amide; a polyvinyl acetate; a polyvinyl acetate copolymer;tylose; a polyvinyl alcohol copolymer; a polyvinylpyrolidone; apolysaccharide, a starch, a modified starch, a starch derivative, adextrin, a maltodextrin, an alginate, a chitosan, a cellulose, acellulose ester, a cellulose ether, a cellulose ether ester,ethylcellulose, methylcellulose, hydroxymethylcellulose,hydroxypropylcellulose, carboxymethylcellulose; a fat; an oil; aprotein, casein, gelatin, zein; a shellac; vinylidene chloride, avinylidene chloride copolymer; a lignosulfonate, calcium lignosulfonate;a polyacrylate, a polymethacrylate, an acrylic copolymer; apolyvinylacrylate; a polyethylene oxide; a polybutene, a polyisobutene,polystyrene, polybutadiene, a polyethyleneamine, a polyethylenamide; anacrylamide polymer or copolymer; a polyhydroxyethyl acrylate, amethylacrylamide; polychloroprene, or any combination thereof.

Embodiment 117 is a composition of any one of embodiments 110-116,wherein the abrading agent comprises talc, graphite, or a combinationthereof.

Embodiment 118 is a composition of any one of embodiments 110-117,wherein the pesticide comprises insecticide, a herbicide, a fungicide, abacteriocide, a nematicide, a miticide, or any combination thereof.

Embodiment 119 is a composition of embodiment 118, wherein theinsecticide comprises clothianidin, imidacloprid, an organophosphate, acarbamate, a pyrethroid, an acaricide, an alkyl phthalate, boric acid, aborate, a fluoride, a sulfur, a haloaromatic substituted urea, ahydrocarbon ester, a biologically-based insecticide, or any combinationthereof.

Embodiment 120 is a composition of embodiment 118, wherein the herbicidecomprises 2,4-D, 2,4-DB, acetochlor, acifluorfen, alachlor, ametryn,atrazine, aminopyralid, benefin, bensulfuron, bensulfuron methyl,bensulide, bentazon, bispyribac sodium, bromacil, bromoxynil, butylate,carfentrazone, chlorimuron, 2-chlorophenoxy acetic acid, chlorsulfuron,chlorimuron ethyl, clethodim, clomazone, clopyralid, cloransulam,CMPP-P_DMA, cycloate, DCPA, desmedipham, dicamba, dichlobenil, diclofop,2,4-dichlorophenol, dichlorophenoxyacetic acid, diclorprop,diclorprop-P, diclosulam, diflufenzopyr, dimethenamid, dimethyl aminesalt of 2,4-dichlorophenoxyacetic acid, diquat, diuron, DSMA, endothall,EPTC, ethalfluralin, ethofumesate, fenoxaprop, fluazifop-P,flucarbazone, flufenacet, flumetsulam, flumiclorac, flumioxazin,fluometuron, fluroxypyr, fluorxypyr 1-methyleptlyester, fomesafen,foresafen sodium salt, foramsulfuron, glufosinate,glufosinate-ammounium, glyphosate, halosulfuron, halosulfuron-methyl,hexazinone, 2-hydroxypehonxy acetic acid, 4-hydroxyphenoxy acetic acid,imazamethabenz, imazamox, imazapic, imazaquin, imazethapyr, isoxaben,isoxaflutole, lactofen, linuron, mazapyr, MCPA, MCPB, mecoprop,mecoprop-P, mesotrione, metolachlor-s, metribuzin, metsulfuron,metsulfuron-methyl, molinate, MSMA, napropamide, naptalam, nicosulfuron,norflurazon, oryzalin, oxadiazon, oxyfluorfen, paraquat, pelargonicacid, pendimethalin, phenmedipham, picloram, prim isulfuron, prodiamine,prometryn, pronamide, propanil, prosulfuron, pyrazon, pyrithiobac,pyroxasulfone, quinclorac, quizalofop, rimsulfuron, sethoxydim, siduron,simazine, sulfentrazone, sulfometuron, sulfosulfuron, tebuthiuron,terbacil, thiazopyr, thifensulfuron, thifensulfuron-methyl, thiobencarb,tralkoxydim, triallate, triasulfuron, tribenuron, tribernuron-methyl,triclopyr, trifluralin, triflusulfuron, or any combination thereof.

Embodiment 121 is a composition of embodiment 118, wherein thenematicide comprises Bacillus firmus, fluopyram, an antibioticnematicide, abamectin, a carbamate nematicide, acetoprole, Bacilluschitonosporus, chloropicrin, benclothiaz, benomyl, Burholderia cepacia,carbofuran, carbosulfan, cleothocard, dazomet, DBCP, DCIP, alanycarb,aldicarb, aldoxycarb, oxamyl, diamidafos, fenamiphos, fosthietan,phosphamidon, cadusafos, chlorpyrifos, diclofenthion, dimethoate,ethoprophos, fensulfothion, fostiazate, harpins, heterophos, imicyafos,isamidofos, isazofos, methomyl, mecarphon, Myrothecium verrucaria,Paecilomyces lilacinus, Pasteuria nishizawae, phorate, phosphocarb,terbufos, thionazin, tioxazafen, triazophos, dazomet,1,2-dicloropropane, 1,3-dichloropropene, furfural, iodomethane, metam,methyl bromide, methyl isothiocyanate, xylenol, or any combinationthereof.

Embodiment 122 is a composition of embodiment 118, wherein thebacteriocide comprises streptomycin, a penicillin, a tetracycline, anoxytetracyline, an ampicillin, an oxolinic acid, kasugamycin,chlorotetracycline, copper oxide or any combination thereof.

Embodiment 123 is a composition of embodiment 122, wherein thebacteriocide comprises oxytetracycline.

Embodiment 124 is a composition of embodiment 118 wherein the biologicalcontrol agent comprises Bacillus thuringiensis, Bacillus megaterium,Bacillus mycoides isolate J, Bacillus methylotrophicus, Bacillusvallismortis, Chromobacterium subtsugae, Delftia acidovorans,Streptomyces lydicus, Streptomyces colombiensis, Streptomyces galbusK61, Penicillium bilaii a lipopeptide-producing Bacillus subtilisstrain, a lipopeptide-producing Bacillus amyloliquefaciens strain, aBacillus firmus strain or a Bacillus pumilus strain.

Embodiment 125 is a composition of any one of embodiments 110-124wherein the fertilizer comprises ammonium sulfate, ammonium nitrate,ammonium sulfate nitrate, ammonium chloride, ammonium bisulfate,ammonium polysulfide, ammonium thiosulfate, aqueous ammonia, anhydrousammonia, ammonium polyphosphate, aluminum sulfate, calcium nitrate,calcium ammonium nitrate, calcium sulfate, calcined magnesite, calciticlimestone, calcium oxide, calcium nitrate, dolomitic limestone, hydratedlime, calcium carbonate, diammonium phosphate, monoammonium phosphate,magnesium nitrate, magnesium sulfate, potassium nitrate, potassiumchloride, potassium magnesium sulfate, potassium sulfate, sodiumnitrates, magnesian limestone, magnesia, urea, urea-formaldehydes, ureaammonium nitrate, sulfur-coated urea, polymer-coated urea, isobutylidenediurea, K₂SO4-Mg₂SO₄, kainite, sylvinite, kieserite, Epsom salts,elemental sulfur, marl, ground oyster shells, fish meal, oil cakes, fishmanure, blood meal, rock phosphate, super phosphates, slag, bone meal,wood ash, manure, bat guano, peat moss, compost, green sand, cottonseedmeal, feather meal, crab meal, fish emulsion, humic acid, or anycombination thereof.

Embodiment 126 is a composition of any one of embodiments 110-125,wherein the fertilizer comprises boric acid, a borate, a boron frit,copper sulfate, a copper frit, a copper chelate, a sodium tetraboratedecahydrate, an iron sulfate, an iron oxide, iron ammonium sulfate, aniron frit, an iron chelate, a manganese sulfate, a manganese oxide, amanganese chelate, a manganese chloride, a manganese frit, a sodiummolybdate, molybdic acid, a zinc sulfate, a zinc oxide, a zinccarbonate, a zinc frit, zinc phosphate, a zinc chelate, or anycombination thereof.

Embodiment 127 is a composition of any one of embodiments 110-126,wherein the biostimulant comprises a seaweed extract, an elicitor, apolysaccharide, a monosaccharide, a protein extract, a soybean extract,a humic acid, a plant hormone, a plant growth regulator, or anycombination thereof.

Embodiment 128 is a composition of any one of embodiments 110-126wherein the amino acid comprises cysteine.

Embodiment 129 is a composition of any one of embodiments or 57-128,further comprising a fungicide.

Embodiment 130 is a composition of embodiment 129, wherein the fungicidecomprises a strobilurin fungicide or a triazole fungicide.

Embodiment 131 is a composition of embodiment 130, wherein thestrobilurin fungicide comprises a Strobilurin A, a Strobilurin B, aStrobilurin C, a Strobilurin D, a Strobilurin E, a Strobilurin F, aStrobilurin G, a Strobilurin H, an Azoxystrobin, a Trifloxystrobin, aKresoxim methyl, a Fluoxastrobin, a Picoxystrobin, or any combinationthereof.

Embodiment 132 is a composition of embodiment 131, wherein thestrobilurin fungicide comprises an Azoxystrobin, a Trifloxystrobin, aKresoxim methyl, a Fluoxastrobin, a Picoxystrobin, a pyraclostrobin, orany combination thereof.

Embodiment 133 is a composition of embodiment 131, wherein thestrobilurin fungicide comprises a Trifloxystrobin.

Embodiment 134 is a composition of embodiment 131, wherein thestrobilurin fungicide comprises a fluoxastrobin.

Embodiment 135 is a composition of any one of embodiments 130-134,wherein the triazole fungicide comprises prothioconazole, imidazole,imidazil, prochloraz, propiconazole, triflumizole, diniconazole,flusilazole, penconazole, hexaconazole, cyproconazole, myclobutanil,tebuconazole, difenoconazole, tetraconazole, fenbuconazole,epoxiconazole, metconazole, fluquinconazole, triticonazole or anycombination thereof.

Embodiment 136 is a composition of any one of embodiments 130-135,wherein the triazole fungicide comprises prothioconazole.

Embodiment 137 is a composition of any one of embodiments 45-52 or57-136, wherein the humectant comprises glycerol, glycerin, a glycerolderivative, glycerol triacetate, triacetin, propylene glycol, hexyleneglycol, butylene glycol, triethylene glycol, tripolypropylene glycol,glyceryl triacetate, sucrose, tagatose, a sugar alcohol, a sugar polyol,sorbitol, xylitol, mannitol, mantitol, a polymeric polyol, polydextrose,collagen, aloe, aloe vera gel, an alpha hydroxy acid, honey, molasses,quillaia, sodium hexametaphosphate, lithium chloride, urea, butyleneglycol, or tremella extract

Embodiment 138 is a composition of any one of embodiments 45-52 or57-137, wherein the carrier comprises water, peat, wheat, bran,vermiculite, clay, pasteurized soil, calcium carbonate, calciumbicarbonate, dolomite, gypsum, bentonite, a clay, a rock phosphate, aphosphorous compound, titanium dioxide, humus, talc, alginate, activatedcharcoal, or a combination thereof.

Embodiment 139 is a composition of any one of embodiments 45-52 or57-138, wherein the composition comprises from about 0.00001 wt. % toabout 95% of the polypeptide(s), from about 0.01% to about 80 wt. % ofthe agrochemical, and from about 5 to about 50 wt % carrier based on thetotal weight of the composition.

Embodiment 140 is a composition of any one of embodiments 45-52 or57-139, wherein the composition comprises from about 0.01% to about 5%of the polypeptide(s), from about 0.2% to about 70 wt. % of theagrochemical, and from about 10 to about 30 wt % carrier based on thetotal weight of the composition.

Embodiment 141 is a composition of any one of embodiments 45-52 or57-139, wherein the composition comprises from about 0.05% to about 1%of the polypeptide(s), from about 30% to about 60 wt. % of theagrochemical, and from about 40 to about 69 wt % carrier based on thetotal weight of the composition.

Embodiment 142 is a composition of any one of embodiments 45-52 or57-141, wherein the agrochemical comprises hydrochloric acid, aceticacid, and/or trifluoroacetic acid.

Embodiment 143 is a composition of embodiment 142 wherein theagrochemical comprises hydrochloric acid, acetic acid, and/ortrifluoroacetic acid and comprises about 0.001 wt. % to about 30% wt. %based on the total weight of the composition.

Embodiment 144 is a composition of embodiment 143 wherein theagrochemical comprises hydrochloric acid, acetic acid, and/ortrifluoroacetic acid and comprises about 0.01 wt. % to about 20% wt. %based on the total weight of the composition.

Embodiment 145 is a composition of embodiment 144 wherein theagrochemical comprises hydrochloric acid, acetic acid, and/ortrifluoroacetic acid and comprises about about 0.1 wt. % to about 5 wt.% based on the total weight of the composition

Embodiment 146 is a seed coated with the polypeptide of any one ofembodiments 1-44 or any combination thereof, or the composition of anyone of embodiments 45-52 and 57-145, or the recombinant microorganism ofany one of embodiments 53-108.

Embodiment 147 is a method for increasing growth, yield, health,longevity, productivity, and/or vigor of a plant or a plant part and/ordecreasing abiotic stress in the plant or the plant part and/orprotecting the plant or the plant part from disease, insects and/ornematodes, and/or increasing the innate immune response of the plant orthe plant part and/or changing plant architecture, the method comprisingeither:

(a) applying the polypeptide of any one of embodiments 1-45 or thecomposition of any one of embodiments 45-52 and 57-145 to a plant, aplant part, or a plant growth medium or a rhizosphere in an areasurrounding the plant or the plant part to increase growth, yield,health, longevity, productivity, and/or vigor of the plant or the plantpart and/or decrease abiotic stress in the plant or the plant partand/or protect the plant or the plant part from disease, insects and/ornematodes, and/or increase the innate immune response of the plant orthe plant part and/or change the plant architecture; or

(b) applying the polypeptide of any one of embodiments 1-45 or thecomposition of any one of embodiments 45-52 and 57-145 to a plant growthmedium to increase growth, yield, health, longevity, productivity,and/or vigor of a plant or a plant part to be grown in the plant growthmedium and/or decrease abiotic stress in the plant or the plant part tobe grown in the plant growth medium and/or protect the plant or theplant part to be grown in the plant growth medium from disease, insectsand/or nematodes, and/or increase the innate immune response and/orchange plant architecture of the plant or the plant part to be grown inthe plant growth medium; or

(c) applying the recombinant microorganism of any one of embodiments53-56, 102 and 105 to a plant, a plant part, or a plant growth medium ora rhizosphere in an area surrounding the plant or the plant part toincrease growth, yield, health, longevity, productivity, and/or vigor ofthe plant or the plant part and/or decrease abiotic stress in the plantor the plant part and/or protect the plant or the plant part fromdisease, insects and/or nematodes, and/or increase the innate immuneresponse of the plant or the plant part and/or change the plantarchitecture, wherein the recombinant microorganism expresses thepolypeptide and expression of the polypeptide is increased as comparedto the expression level the polypeptide in a wild-type microorganism ofthe same kind under the same conditions.

Embodiment 148 is a method of embodiment 147, wherein the polypeptide orthe composition comprises: the Flg22 polypeptide and an amino acidsequence of the Flg22 polypeptide comprises any one of SEQ ID NOs:226-300 and 571-573 or any combination thereof; the retro inverso Flg22polypeptide and an amino acid sequence of the retro inverso Flg22polypeptide comprises any one of SEQ ID NO: 376-450 or any combinationthereof; or any combination thereof to decrease abiotic stress in theplant or the plant part and/or protect the plant or the plant part fromdisease and/or increase the innate immune response of the plant or theplant part.

Embodiment 149 is a method of embodiment 147, wherein the polypeptide orthe composition comprises: the FlgII-28 polypeptide and an amino acidsequence of the FlgII-28 polypeptide comprises any one of SEQ ID NOs:301-375 or any combination thereof; the retro inverso FlgII-28polypeptide and an amino acid sequence of the retro inverso FlgII-28polypeptide comprises any one of SEQ ID NO: 451-525 or any combinationthereof; or any combination thereof to decrease abiotic stress in theplant or the plant part and/or protect the plant or the plant part fromdisease and/or increase the innate immune response of the plant or theplant part.

Embodiment 150 is a method of embodiment 147, wherein the polypeptide orthe composition comprises: the retro inverso Flg22 polypeptide and anamino acid sequence of the retro inverso Flg22 polypeptide comprises anyone of SEQ ID NO: 376-450 or any combination thereof; the retro inversoFlgII-28 polypeptide and an amino acid sequence of the retro inversoFlgII-28 polypeptide comprises any one of SEQ ID NO: 451-525 or anycombination thereof; or any combination thereof to decrease abioticstress in the plant or the plant part and/or protect the plant or theplant part from disease and/or increase the innate immune response ofthe plant or the plant part.

Embodiment 151 is a method of embodiment 147, wherein the polypeptide orthe composition comprises: the RHPP polypeptide and an amino acidsequence of the RHPP polypeptide comprises SEQ ID NO: 600, 603, 604 orany combination thereof; the Kunitz Trypsin Inhibitor (KTI) polypeptideand an amino acid sequence of the KTI polypeptide comprises SEQ ID NO:602; the retro-inverso RHPP polypeptide and an amino acid sequence ofthe RI RHPP comprises SEQ ID NO 601, 605, 606 or any combinationthereof; or any combination thereof to decrease abiotic stress in theplant or the plant part and/or protect the plant or the plant part fromdisease and/or increase the innate immune response of the plant or theplant part.

Embodiment 152 is a method of any one of embodiments 147 to 151, whereinthe abiotic stress comprises heat stress, temperature stress, radiationstress, drought stress, cold stress, salt stress, nutrient-deficientstress, high metal stress, water stress, osmotic stress, or anycombination thereof.

Embodiment 153 is a method of embodiment 147, wherein the polypeptide orthe composition comprises: the Flg22 polypeptide and an amino acidsequence of the Flg22 polypeptide comprises any one of SEQ ID NOs:226-300 and 571-573; the retro inverso Flg22 polypeptide and an aminoacid sequence of the retro inverso Flg22 polypeptide comprises any oneof SEQ ID NO: 376-450; or any combination thereof to protect the plantor the plant part from disease and/or increase the innate immuneresponse of the plant or the plant part.

Embodiment 154 is a method of embodiment 147 or 153, wherein thepolypeptide or the composition comprises: the FlgII-28 polypeptide andan amino acid sequence of the FlgII-28 polypeptide comprises any one ofSEQ ID NOs: 301-375; the retro inverso FlgII-28 polypeptide and an aminoacid sequence of the retro inverso FlgII-28 polypeptide comprises anyone of SEQ ID NO: 451-525; or any combination thereof to protect theplant or the plant part from disease and/or increase the innate immuneresponse of the plant or the plant part.

Embodiment 155 is a method of embodiment 147 wherein the polypeptide orthe composition comprises: the Flg22 polypeptide and an amino acidsequence of the Flg22 polypeptide comprises SEQ ID NO: 226, 752 or 571and EF-Tu polypeptides and the amino acid sequences of the EF-Tupolypeptides comprise SEQ ID NOs 616 and 617 to protect the plant or theplant part from disease and/or increase the innate immune response ofthe plant or the plant part.

Embodiment 156 is a method of embodiment 147, wherein the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 289, 290, 291, 293, 294, 295, 300, 437, 532,534, 536, 538, 540, 571-586, and 751-766 or any combination thereof toprotect the plant or the plant part from disease, insects or nematodes.

Embodiment 157 is a method of embodiment 156, wherein the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 293, 295, 300, 540, 571 574, 751, and 752 orany combination thereof.

Embodiment 158 is a method of any one of embodiments 147 and 150-157,wherein the disease comprises Asian citrus greening, Huanglonging (HLB)disease, Asian soybean rust, Sclerotinia stem rot (or white mold),Pseudomonas leaf spot, or Cercospora leaf blight.

Embodiment 159 is a method of embodiment 158, wherein the polypeptide orthe composition comprises the Flg22 polypeptide and an amino acidsequence of the Flg22 polypeptide comprises any one of SEQ ID NOs:226-300 and 571-573 or any combination thereof.

Embodiment 160 is a method of embodiment 158, wherein the polypeptide orthe composition comprises the FlgII-28 polypeptide and an amino acidsequence of the FlgII-28 polypeptide comprises any one of SEQ ID NOs:301-375, or 751 or any combination thereof.

Embodiment 161 is a method of embodiment 158, wherein the polypeptide orthe composition comprises the Flg22 polypeptide and the FlgII-28polypeptide, an amino acid sequence of the Flg22 polypeptide comprisingany one of SEQ ID NOs: 226-300 and 571-573 or any combination thereofand an amino acid sequence of the FlgII-28 polypeptide comprising anyone of SEQ ID NOs: 301-375 or 751 or any combination thereof.

Embodiment 162 is a method of embodiment 161, wherein the polypeptide orthe composition further comprises the retro inverso Flg22 polypeptide,the retro inverso FlgII-28 polypeptide or a combination thereof, anamino acid sequence of the retro inverso Flg22 polypeptide comprisingany one of SEQ ID NO: 376-450 or any combination thereof and an aminoacid sequence of the retro inverso FlgII-28 polypeptide comprising anyone of SEQ ID NO: 451-525 or any combination thereof.

Embodiment 163 is a method of embodiment 147, wherein the polypeptide orthe composition comprises the RHPP polypeptide and/or the RI RHPPpolypeptide to increase the yield, the growth and/or the productivity ofthe plant or plant part and/or change the plant architecture.

Embodiment 164 is a method of embodiment 163, wherein the RHPPpolypeptide comprises SEQ ID NO: 600.

Embodiment 165 is a method of embodiment 163 or 164, wherein the growthcomprises root growth, root length, root biomass, nodulation, totalbiomass, above ground biomass, or any combination thereof.

Embodiment 166 is a method of any one of embodiments 163-165, whereinthe plant comprises soybean, the growth comprises overall root length,root biomass, nodulation, nodules per plant, total biomass, above groundbiomass, or any combination thereof, and the productivity comprisesnumber of total pods or pods per node.

Embodiment 167 is a method of any one of embodiments 163-166, whereinthe plant architecture comprises beneficial outcomes to the plant orplant part.

Embodiment 168 is a method of embodiment 167, wherein the beneficialoutcomes comprise increased planting density capability for a field ofthe plants.

Embodiment 169 is a method of embodiment 147, wherein the polypeptide orthe composition comprises the harpin-like polypeptide or the RHPPpolypeptide to protect the plant or the plant part from disease, insectsand/or nematodes, and/or increase the innate immune response of theplant or the plant part.

Embodiment 170 is a method of embodiment 147, wherein the polypeptide orthe composition comprises the PSK polypeptide to increase yield of theplant or the plant part in environments prone to heat and drought.

Embodiment 171 is a method of embodiment 170, wherein the polypeptide,the composition, or the recombinant microorganism is applied just priorto floral formation or at the pre-flowering stage.

Embodiment 172 is a method of embodiment 147, wherein the polypeptide orthe composition comprises the PSK polypeptide, the RHPP, the harpin orharpin-like polypeptide, or a combination thereof to increase growth ofthe plant or the plant part.

Embodiment 173 is a method of embodiment 172, wherein the growthcomprises root and floral apical meristems, floral organ production,fruit development, fruit production, number of floral organs, size offloral organs, or a combination thereof.

Embodiment 174 is a method of embodiment 147, wherein the polypeptide orthe composition comprises the PSK polypeptide and the harpin orharpin-like polypeptide to increase growth and productivity of the plantor the plant part in an environment prone to both stress and non-stressconditions for plant growth.

Embodiment 175 is a method of embodiment 147, wherein the polypeptide orthe composition comprises the thionin or thionin-like polypeptide.

Embodiment 176 is a method of embodiment 175, wherein the thionin orthionin-like polypeptide is fused to a phloem targeting sequence to forma fused polypeptide, the amino acid sequence of the phloem targetingsequence comprising any one of SEQ ID NOs: 641-649, or any combinationthereof, for delivering the fused polypeptide to vascular tissue orcells and/or phloem or phloem-associated tissue or cells in the plant orplant part.

Embodiment 177 is a method of embodiment 147, wherein the polypeptidescomprise Flg22 polypeptides.

Embodiment 178 is a method of embodiment 177, wherein the polypeptidescomprise SEQ ID NOs 526 and 527.

Embodiment 179 is a method of embodiment 177, wherein the polypeptidescomprise SEQ ID NOs 226 and 227.

Embodiment 180 is a method of embodiment 175 or 176, wherein the diseasecomprises Asian Citrus disease (HLB), Citrus Canker disease, Cercosporaleaf blight, or a bacteria causing disease.

Embodiment 181 is a method of embodiment 180, wherein the bacteriacausing disease comprises bacterial leaf blight, bacterial stalk rot,bacterial leaf spot, bacterial leaf scorch, bacterial top rot, bacterialstripe, chocolate spot, Goss's bacterial wilt and blight, Holcus spot,purple leaf sheath, seed rot, seedling blight, Stewart's disease(bacterial wilt), corn stunt, Fire Blight, Pierce's disease, citrusvariegated chlorosis, citrus canker, Pseudomonas syringae serovars, or acombination thereof.

Embodiment 182 is a method of embodiment 147, wherein the polypeptide orthe composition comprises the flagellin or flagellin-like polypeptide,and an amino acid sequence of the flagellin or flagellin-likepolypeptide comprising any one of SEQ ID NOs: 226-525 and 571-573 or anycombination thereof.

Embodiment 183 is a method of embodiment 147 wherein the diseasecomprises Asian soybean rust, Cercospora or Sclerotinia stem rot (orwhite mold) and the polypeptide or composition comprises:

a flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 752, and 571; or

an RHPP polypeptide and the amino acid sequence of the RHPP polypeptidecomprises SEQ ID NO: 600; or

a flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 752, and 571 and an RHPP polypeptide and theamino acid sequence of the RHPP polypeptide comprises SEQ ID NO: 600.

Embodiment 184 is a method of embodiment 183 wherein the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226.

Embodiment 185 is a method of 183 or 184 wherein the composition furthercomprises a fungicide.

Embodiment 186 is a method of embodiment 147 wherein the diseasecomprises a bacterial disease, and the method comprises restrictinggrowth of a bacteria and/or preventing the disease and wherein the aminoacid sequence of the flagellin- or flagellin associated polypeptidecomprises any one of SEQ ID NOs: 226, 540, 752, and 571 or anycombination thereof.

Embodiment 187 is a method of embodiment 186 wherein the bacteriacomprises Pseudomonas syringae pv. actinidiae.

Embodiment 188 is a method of embodiment 187 wherein the plant comprisesa kiwi plant.

Embodiment 189 is a method of embodiment 147 wherein the diseasecomprises Asian citrus greening and the composition comprises abacteriocide and the flagellin or flagellin-associated polypeptide andthe amino acid sequence of the flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 226, 571, and 752 or anycombination thereof.

Embodiment 190 is a method of 189 wherein the bacteriocide comprisesoxytetracycline.

Embodiment 191 is a method of embodiment 147 wherein the diseasecomprises Sclerotinia stem rot (or white mold) and the compositioncomprises the flagellin or flagellin-associated polypeptide and theamino acid sequence of the flagellin or flagellin-associated polypeptidecomprises any one of SEQ ID NOs: 226, 540, 571, 751 and 752.

Embodiment 192 is a method of any one of embodiments 147-191, whereinprotecting the plant or the plant part from disease comprisesprophylactic treatment, treatment, prevention and decreased diseaseprogression on or in the plant or plant part.

Embodiment 193 is a method of any one of embodiments 147-192, whereinthe polypeptide, the composition, or the recombinant microorganism isapplied exogenously to the plant, the plant part, or the plant growthmedium.

Embodiment 194 is a method of any one of embodiments 147-193, whereinthe polypeptide, the composition, or the recombinant microorganism isapplied endogenously to the plant or the plant part.

Embodiment 195 is a method of any one of embodiments 147-194, whereinthe plant part comprises a cell, a leaf, a branch, a stem, a flower, afoliage, a floral organ, a fruit, pollen, a vegetable, a tuber, a corm,a bulb, a pseudobulb, a pod, a root, a root ball, a root stock, a scion,or a seed.

Embodiment 196 is a method of any one of embodiments 147-195, whereinthe polypeptide, the composition, or the recombinant microorganism isapplied to a surface of the plant, a foliage of the plant or a surfaceof a seed of the plant.

Embodiment 197 is a method of embodiment 196, wherein the polypeptide,the composition, or the recombinant microorganism is applied to thesurface of the seed and the plant or the plant part is grown from theseed.

Embodiment 198 is a method of any one of embodiments 147-197, whereinthe polypeptide, the composition, or the recombinant microorganism isapplied as a foliar application.

Embodiment 199 is a method of any one of embodiments 147-198, whereinthe plant is a fruit plant or a vegetable plant, and the method providesincreased yield of fruits or vegetables.

Embodiment 200 is a method of producing a polypeptide, comprisingproducing a fusion protein comprising a polypeptide of any one ofEmbodiments 1 to 44, and an enterokinase (EK) cleavage site viafermentation, the enterokinase cleavage site enhancing activity andstability of the polypeptide.

Embodiment 201 is a method of embodiment 200, further comprisingapplying an enterokinase to the prepared fusion protein to cleave theenterokinase cleavage site and isolate the polypeptide.

Embodiment 202 is a method of embodiment 200 or 201, wherein theenterokinase cleavage site comprises SEQ ID NO: 772.

Embodiment 203 is a method of any one of embodiments 200 to 202, whereinthe fusion protein further comprises a protein tag.

Embodiment 204 is a method of embodiment 203, wherein the protein tagcomprises glutathione S transferase (GST).

Embodiment 205 is the method of any one of embodiments 200 to 204,wherein the fusion protein further comprises a secretion signal.

Embodiment 206 is the method of embodiment 205, wherein the secretionsignal comprises an amino acid sequence comprising any one of SEQ IDNOs: 563-570, or 769, or any combination thereof.

What is claimed is:
 1. A polypeptide for bioactive priming of a plant ora plant part to increase growth, yield, health, longevity, productivity,and/or vigor of a plant or a plant part and/or decrease abiotic stressin the plant or the plant part and/or protect the plant or the plantpart from disease, insects and/or nematodes, and/or increase the innateimmune response of the plant or the plant part and/or change plantarchitecture, wherein the polypeptide comprises: (a) a flagellin orflagellin-associated polypeptide and an amino acid sequence of theflagellin or flagellin-associated polypeptide comprises any one of SEQID NOs: 226, 1-225, 227-375, 526, 528, 530, 532, 534, 536, 538, 540,541, 751, 752, and 754-766; or (b) a mutant flagellin orflagellin-associated polypeptide and an amino acid sequence of themutant flagellin or flagellin-associated polypeptide comprises any oneof SEQ ID NOs: 571-579, and 753; or (c) a mutant flagellin orflagellin-associated polypeptide and an amino acid sequence of themutant flagellin or flagellin-associated polypeptide comprises any oneof SEQ ID NOs: 580-586; or (d) a retro inverso Flg22 polypeptide and anamino acid sequence of the retro inverso Flg22 polypeptide comprises anyone of SEQ ID NOs: 376-450, 527, 531, 533, 535, 537 and 539; or (e) aretro inverso FlgII-28 polypeptide and an amino acid sequence of theretro inverso FlgII-28 polypeptide comprises any one of SEQ ID NOs:451-525, or 768; or (f) a retro inverso Flg15 polypeptide and an aminoacid sequence of the retro inverso Flg15 polypeptide comprises SEQ IDNOs: 529 or 767; or (g) a harpin or harpin-like polypeptide and an aminoacid sequence of the harpin or harpin-like polypeptide comprises any oneof SEQ ID NOs: 587, 589, 591, 593, 594 and 595; or (h) a retro inversoharpin or harpin-like polypeptide and an amino acid sequence of theretro inverso harpin or harpin-like polypeptide comprises any one of SEQID NOs: 588, 590, 592, 596 and 597; or (i) a root hair promotingpolypeptide (RHPP) and an amino acid sequence of the RHPP comprises anyone of SEQ ID Nos: 600, 603 and 604; or (j) a Kunitz Trypsin Inhibitor(KTI) polypeptide and an amino acid sequence of the KTI polypeptidecomprises SEQ ID No: 602; or (k) a retro inverso root hair promotingpolypeptide (RI RHPP) and an amino acid sequence of the RI RHPPcomprises any one of SEQ ID NO: 601, 605 and 606; or (l) an elongationfactor Tu (EF-Tu) polypeptide and an amino acid sequence of the EF-Tupolypeptide comprises any one of SEQ ID NOs: 607-623; or (m) a retroinverso elongation factor Tu (RI EF-Tu) polypeptide and an amino acidsequence of the RI EF-Tu polypeptide comprises any one of SEQ ID NOs:624-640; or (n) a fusion polypeptide comprising SEQ ID NO: 750; or (o) aphytosulfokine (PSK) polypeptide and an amino acid sequence of the PSKpolypeptide comprises SEQ ID NO: 598; or (p) a retro inversophytosulfokine (RI PSK) polypeptide and an amino acid sequence of the RIPSK polypeptide comprises SEQ ID NO: 599; or (q) a thionin orthionin-like polypeptide and an amino acid sequence of the thionin orthionin-like polypeptide comprises any one of SEQ ID NOs: 650-749, andoptionally, wherein the flagellin or flagellin-associated polypeptide of(a), the mutant flagellin or flagellin-associated polypeptide of (c),the harpin or harpin-like polypeptide of (g), the KTI polypeptide of(j), the PSK polypeptide of (o), and the thionin or thionin-likepolypeptide of (q) either: contains a chemical modification; is avariant having an amino acid insertion, deletion, inversion, repeat,duplication, extension, or substitution within the amino acid; or ispart of a fusion protein; or contains a protease recognition sequence orfurther comprises a core sequence.
 2. The polypeptide of claim 1,wherein the flagellin or flagellin-associated polypeptide of (a), themutant flagellin or flagellin-associated polypeptide of (c), the harpinor harpin-like polypeptide of (g), the KTI polypeptide of (j), the PSKpolypeptide of (o), and the thionin or thionin-like polypeptide of (q)either: contains a chemical modification; is a variant having an aminoacid insertion, deletion, inversion, repeat, duplication, extension, orsubstitution within the amino acid; or is part of a fusion protein; orcontains a protease recognition sequence; or further comprises a coresequence.
 3. The polypeptide of claim 2, wherein the chemicalmodification comprises acetylation, acid addition, acylation,ADP-ribosylation, aldehyde addition, alkylamide addition, amidation,amination, biotinylation, carbamate addition, chloromethyl ketoneaddition, covalent attachment of a nucleotide or nucleotide derivative,cross-linking, cyclization, disulfide bond formation, demethylation,ester addition, formation of covalent cross-links, formation ofcysteine-cysteine disulfide bonds, formation of pyroglutamate,formylation, gamma-carboxylation, glycosylation, GPI anchor formation,hydrazide addition, hydroxyamic acid addition, hydroxylation,iodination, lipid addition, methylation, myristoylation, oxidation,PEGylation, proteolytic processing, phosphorylation, prenylation,palmitoylation, addition of a purification tag, pyroglutamyl addition,racemization, selenoylation, sulfonamide addition, sulfation,transfer-RNA mediated addition of amino acids to proteins,ubiquitination, urea addition, or an N-terminal modification or aC-terminal modification; or the amino acid substitution within the aminoacid of the variant comprises a substitution of a β-amino acid, aD-amino acid, or a non-natural amino acid.
 4. The polypeptide of claim3, wherein the chemical modification comprises acetylation, amidation,cross-linking, or cyclization.
 5. The polypeptide of claim 2, wherein:the amino acid sequence of the flagellin or flagellin associatedpolypeptide comprises any one of SEQ ID NOs 754-766; or the mutantflagellin or flagellin associated polypeptide is from a Bacillus, aLysinibacillus, a Paenibacillus, or an Aneurinbacillus genus bacterium.6. The polypeptide of claim 1, wherein the polypeptide comprises: themutant flagellin or flagellin-associated polypeptide of (b); or theretro inverso Flg22 polypeptide of (d); or the retro inverso FlgII-28polypeptide of (e); or the retro inverso Flg15 polypeptide of (f); orthe retro inverso harpin or harpin-like polypeptide of (h); or the RHPPof (i); or the RI RHPP of (k); or the EF-Tu polypeptide of (l); or theRI EF-Tu polypeptide of (m); or the fusion polypeptide of (n); the RIPSK polypeptide of (p); or a core sequence.
 7. The polypeptide of claim6 wherein the mutant flagellin or flagellin associated polypeptide of(b) comprises any one of SEQ ID NOs 571-573; and/or the core sequencecomprises any one of SEQ ID NOs 754-766; and/or the fusion polypeptidecomprises an assistance polypeptide.
 8. The polypeptide of claim 7wherein the assistance polypeptide comprises: a signature polypeptide,and an amino acid sequence of the signature polypeptide comprises anyone of SEQ ID NOs: 542-548, or any combination thereof; or a signalanchor sorting polypeptide, and an amino acid sequence of the signalanchor sorting polypeptide comprises any one of SEQ ID NOs: 549-562, orany combination thereof; or a signature polypeptide and an amino acidsequence of the signature polypeptide comprises SEQ ID NO: 542; or asignal anchor sorting polypeptide and an amino acid sequence of thesignal anchor sorting polypeptide comprises SEQ ID NO:
 549. 9. Thepolypeptide of any one of claims 1 to 8 wherein the polypeptide furthercomprises the core sequence comprising any one of SEQ ID NOs: 754-766and wherein the inclusion of the core sequence increases the bioactivepriming activity of the polypeptide.
 10. The polypeptide of any one ofclaims 1 to 9 wherein the polypeptide comprises an amino acid sequencehaving at least 70%, at least 75%, at least 80%, at least 85%, at least90%, at least 95%, at least 98% or at least 99% identity to any one ofSEQ ID NOs. 1-768, and the polypeptide has bioactive priming activity.11. The polypeptide of any one of claims 1 to 10 wherein the polypeptideis isolated, concentrated from a fermentation product, and/or partiallypurified, optionally, by filtration, chromatography or from arecombinant microorganism.
 12. A composition for bioactive priming of aplant or a plant part to increase growth, yield, health, longevity,productivity, and/or vigor of a plant or a plant part and/or decreaseabiotic stress in the plant or the plant part and/or protect the plantor the plant part from disease, insects and/or nematodes, and/orincrease the innate immune response of the plant or the plant partand/or change plant architecture, the composition comprising either: thepolypeptide of any one of claims 1-11, or any combination thereof, andan agrochemical or a carrier; or any combination of the polypeptides ofany one of claims 1-12.
 13. The composition of claim 12, wherein thecomposition comprises: (A) the flagellin or flagellin-associatedpolypeptide of (a), the mutant flagellin or flagellin-associatedpolypeptide of (c), the harpin or harpin-like polypeptide of (g), theKTI polypeptide of (j), the PSK polypeptide of (o), the thionin orthionin-like polypeptide of (q), or any combination thereof, and anagrochemical or carrier, the agrochemical or carrier not beingassociated with the polypeptide in nature; or (B) the mutant flagellinor flagellin-associated polypeptide of (b); or the retro inverso Flg22polypeptide of (d); or the retro inverso FlgII-28 polypeptide of (e); orthe retro inverso Flg15 polypeptide of (f); or the retro inverso harpinor harpin-like polypeptide of (h); or the RHPP of (i); or the RI RHPP of(k); or the EF-Tu polypeptide of (l); or the RI EF-Tu polypeptide of(m); or the fusion polypeptide of (n); or the RI PSK polypeptide of (p),or any combination thereof; and the flagellin or flagellin-associatedpolypeptide of (a), the mutant flagellin or flagellin-associatedpolypeptide of (c), the harpin or harpin-like polypeptide of (g), theKTI polypeptide of (j), the PSK polypeptide of (o), the thionin orthionin-like polypeptide of (q), or any combination thereof; or (C) twoor more of the mutant flagellin or flagellin-associated polypeptide of(b); or the retro inverso Flg22 polypeptide of (d); or the retro inversoFlgII-28 polypeptide of (e); or the retro inverso Flg15 polypeptide of(f); or the retro inverso harpin or harpin-like polypeptide of (h); orthe RHPP of (i); or the RI RHPP of (k); or the EF-Tu polypeptide of (l);or the RI EF-Tu polypeptide of (m); or the fusion polypeptide of (n); orthe RI PSK polypeptide of (p); or (D) two or more of the flagellin orflagellin-associated polypeptide of (a), the mutant flagellin orflagellin-associated polypeptide of (c), the harpin or harpin-likepolypeptide of (g), the KTI polypeptide of (j), the PSK polypeptide of(o), or the thionin or thionin-like polypeptide of (q), wherein either:(i) at least one of the flagellin or flagellin-associated polypeptide of(a), the mutant flagellin or flagellin-associated polypeptide of (c),the harpin or harpin-like polypeptide of (g), the PSK polypeptide of(o), or the thionin or thionin-like polypeptide of (q) contains thechemical modification; is the variant; is part of the fusion protein; orcontains the protease recognition sequence; or (ii) at least two of theflagellin or flagellin-associated polypeptide of (a), the mutantflagellin or flagellin-associated polypeptide of (c), the harpin orharpin-like polypeptide of (g), the KTI polypeptide of (j), the PSKpolypeptide of (o), or the thionin or thionin-like polypeptide of (q)are not associated in nature; or (E) wherein the flagellin orflagellin-associated polypeptide is produced recombinantly by amicroorganism.
 14. The composition of claim 13 wherein the compositioncomprises: (i) the composition of (A) or the composition of (B) andwherein the flagellin or flagellin-associated polypeptide of (a), themutant flagellin or flagellin-associated polypeptide of (c), the harpinor harpin-like polypeptide of (g), the KTI polypeptide of (j), the PSKpolypeptide of (o), the thionin or thionin-like polypeptide of (q), orany combination thereof does not contain the chemical modification; isnot the variant; is not part of the fusion protein; and does not containthe protease recognition sequence; or (ii) the composition of (E)wherein the microorganism comprises a bacterium from the genus Bacillus,a bacterium from the genus Pseudomonas, a bacterium from the genusPaenibacillus, a fungus of the genus Penicillium, a bacterium of thegenus Glomus, a bacterium of the genus Arthrobacter, a bacterium of thegenus Paracoccus, a bacterium of the genus Rhizobium, a bacterium of thegenus Bradyrhizobium, a bacterium of the genus Azosprillium, a bacteriumof the genus Enterobacter, a bacterium of the genus Escherichia, or anycombination thereof.
 15. A recombinant microorganism that expresses oroverexpresses a polypeptide, wherein the polypeptide comprises thepolypeptide of any one of claims 1-15 or any combination thereof. 16.The recombinant microorganism of claim 15, wherein the polypeptidecomprises: the flagellin or flagellin-associated polypeptide of (a); orthe mutant flagellin or flagellin-associated polypeptide of (b); or themutant flagellin or flagellin-associated polypeptide of (c); or theharpin or harpin-like polypeptide of (g); or the RHPP of (i); or the KTIpolypeptide of (j); or the EF-Tu polypeptide of (l); or the fusionpolypeptide of (n); or the PSK polypeptide of (o); or the thionin orthionin-like polypeptide of (q); and/or the polypeptide is overexpressedby the microorganism.
 17. The recombinant microorganism of claim 16,wherein the polypeptide is overexpressed by the microorganism and ismade with a secretion signal.
 18. The composition or recombinantmicroorganism of any one of claims 12-17 wherein the polypeptidecomprises: the flagellin or flagellin associated polypeptide; the retroinverso Flg22 polypeptide; the retro inverso FlgII-28 polypeptide; thethionin or thionin-like polypeptide; the thionin or thionin-likepolypeptide wherein the amino acid sequence of the thionin orthionin-like polypeptide comprises SEQ ID NO: 651; the PSK polypeptide;the PSK polypeptide and the amino acid sequence of the PSK polypeptidecomprises SEQ ID NO: 598; the RHPP; the RHPP and the amino acid sequenceof the RHPP comprises SEQ ID NO: 600; the RI RHPP; the harpin orharpin-like polypeptide; the harpin or harpin-like polypeptide and theamino acid sequence of the harpin or harpin-like polypeptide comprisesSEQ ID NO: 587; the EF-Tu polypeptide; or the fusion polypeptide; or thecomposition or recombinant microorganism comprises the PSK polypeptide,the RHPP, the harpin or harpin-like polypeptide, or a combinationthereof; or the flagellin or flagellin-associated polypeptide is afusion protein with a polypeptide; or the polypeptide further comprisesa secretion signal.
 19. The composition of claim 18 wherein thepolypeptide comprises the flagellin or flagellin-associated polypeptideand, wherein: (i) flagellin or flagellin-associated polypeptide ismodified chemically on its N or C terminus; or (ii) the flagellin orflagellin-associated polypeptide is modified via crosslinking orcyclization; or (iii) the flagellin or flagellin-associated polypeptideis from a Bacillus, a Lysinibacillus, a Paenibacillus, anAneurinibacillus genus bacterium, or any combination thereof; or (iv)the amino acid sequence of the flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 1-75, or any combinationthereof; or (v) the flagellin or flagellin-associated polypeptidecomprises a truncated N-terminal polypeptide and an amino acid sequenceof the truncated N-terminal polypeptide comprises SEQ ID NO: 76, 78, 80,82, 84, 86, 88, 90, 92, 94, 96, 98, 100, 102, 104, 106, 108, 109, 110,112, 114, 116, 118, 120, 122, 124, 126, 128, 130, 132, 134, 136, 138,140, 142, 144, 146, 148, 150, 152, 154, 156, 158, 160, 162, 164, 166,168, 170, 172, 174, 176, 178, 180, 182, 184, 186, 188, 190, 192, 194,196, 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222,224, 752, or any combination thereof; or (vi) the flagellin orflagellin-associated polypeptide comprises a truncated C-terminalpolypeptide and an amino acid sequence of the truncated C-terminalpolypeptide comprises SEQ ID NO: 77, 79, 81, 83, 85, 87, 89, 91, 93, 95,97, 99, 101, 103, 105, 107, 109, 111, 113, 115, 117, 119, 121, 123, 125,127, 129, 131, 133, 135, 137, 141, 143, 145, 147, 149, 151, 153, 155,157, 159, 161, 163, 165, 167, 169, 171, 173, 175, 177, 179, 181, 183,185, 187, 189, 191, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219,221, 223, 225, or any combination thereof; or (vii) the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 289, 290, 291, 293, 294, 295, 300, 437, 532,534, 536, 538, 540, 571-586, and 751-768 or any combination thereof; or(viii) the amino acid sequence of the flagellin or flagellin-associatedpolypeptide comprises any one of SEQ ID NOs: 226-300, or any combinationthereof; or (ix) the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs:301-375, and 751 or any combination thereof
 20. The composition orrecombinant microorganism of claim 19 wherein the amino acid sequence ofthe flagellin or flagellin associated polypeptide comprises: any one ofSEQ ID NOs: 226, 293, 295, 300, 540, 571, 574 and 752, or anycombination thereof; or any one of SEQ ID NOs 754-766; or any one of SEQID NOs 754-766 wherein the flagellin or flagellin-associated polypeptideis incorporated into a non-flagellin or flagellin-associated polypeptideor full length protein, wherein the incorporation of the flagellin orflagellin-associated polypeptide results in increased bioactive primingactivity in the non-flagellin or flagellin-associated polypeptide orfull length protein; or SEQ ID NO:
 301. 21. The composition orrecombinant microorganism of claim 18 wherein: (i) the thionin orthionin-like polypeptide is fused to a phloem targeting sequence to forma fused polypeptide, the amino acid sequence of the phloem targetingsequence comprising any one of SEQ ID NOs: 641-649, or any combinationthereof, for delivering the fused polypeptide to vascular tissue orcells and/or phloem or phloem-associated tissue or cells in the plant orplant part; or (ii) the thionin or thionin-like polypeptide wherein theamino acid sequence of the thionin or thionin-like polypeptide comprisesSEQ ID NO: 651 and the thionin or thionin-like polypeptide is fused to aphloem targeting sequence to form a fused polypeptide, the amino acidsequence of the phloem targeting sequence comprising any one of SEQ IDNO: 641, or any combination thereof, for delivering the fusedpolypeptide to vascular tissue or cells and/or phloem orphloem-associated tissue or cells in the plant or plant part; or (iii)the amino acid sequence of the RHPP comprises SEQ ID NO: 600 and thecomposition or recombinant microorganism further comprising theflagellin or flagellin-associate polypeptide; or (iv) the amino acidsequence of the RHPP comprises SEQ ID NO: 600 and the composition orrecombinant microorganism further comprising the flagellin orflagellin-associate polypeptide and the amino acid sequence of theflagellin or flagellin-associated polypeptide comprises any one of SEQID NOs: 226, 752, and 571; (v) the amino acid sequence of the RHPPcomprises SEQ ID NO: 600 and the composition or recombinantmicroorganism further comprising the flagellin or flagellin-associatepolypeptide and the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises SEQ ID NO: 226; or (vi) theamino acid sequence of the EF-Tu polypeptide comprises SEQ ID NO: 616;or (vii) the polypeptide further comprises the secretion signal and theamino acid sequence of the EF-Tu polypeptide comprises SEQ ID NO: 616;or (viii) the polypeptide further comprises the secretion signal and theamino acid sequence of the EF-Tu polypeptides comprise SEQ ID NOs: 616and
 617. 22. The composition or recombinant microorganism of claim 18 or21 wherein the composition or recombinant microorganism comprises theEF-Tu polypeptide or the EF-Tu polypeptide and the secretion signal, andfurther comprises the flagellin or flagellin-associated polypeptide, theamino acid sequence of the flagellin or flagellin-associated polypeptidecomprising any one of SEQ ID NOs: 1-525, 526, 530, 532, 534, 536, 538,540, 541, 571-586, and 751-766, or any combination thereof.
 23. Thecomposition or recombinant microorganism of claim 22 wherein the aminoacid sequence of the flagellin or flagellin-associated polypeptidecomprising SEQ ID NO: 226, 571 or 752, or a combination thereof; or thecomposition or recombinant microorganism further comprises the harpin orharpin-like polypeptide; or the composition or recombinant microorganismfurther comprises the harpin or harpin-like polypeptide and the harpinor harpin-like polypeptide comprises SEQ ID NO:
 587. 24. The compositionor recombinant microorganism of any one of claims 12-17, comprising (a)the flagellin or flagellin-associated polypeptides and the amino acidsequences of the flagellin or flagellin-associated polypeptides compriseSEQ ID NOs: 571, 295, 300, 293, and 580; or 295, 300, 293, and 580; or571, 295, 293, and 580; or 571, 300, 293, and 580; or 571, 293 and 580;or 571, 295, 293; or (b) the flagellin or flagellin-associatedpolypeptide and the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises SEQ ID NO: 226 and one ormore of cellobiose, cellulose, chitin, or chitosan; or (c) the flagellinor flagellin-associated polypeptide and the amino acid sequence of theflagellin or flagellin-associated polypeptide comprises SEQ ID NO: 226and the harpin or harpin-like polypeptide and the amino acid sequence ofthe harpin or harpin-like polypeptide comprises SEQ ID NO: 591; or (d)the harpin or harpin-like polypeptide and the amino acid sequence of theharpin or harpin-like polypeptide comprises SEQ ID NO: 587 and the PSKpolypeptide and the amino acid sequence of the PSK polypeptide comprisesSEQ ID NO: 598; or (e) the flagellin or flagellin-associated polypeptideand the amino acid sequence—of the flagellin or flagellin-associatedpolypeptide—comprises SEQ ID NO: 226, 571 or 752 or any combinationthereof and the EF-Tu polypeptides and the amino acid sequences of theEF-Tu polypeptides comprise SEQ ID NOs: 616 and 617; or (f) theflagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226, 540, 571 or 752, or any combination thereof; or (g) theRHPP polypeptide and the amino acid sequence of the RHPP polypeptidecomprises SEQ ID NO: 600; or (h) the flagellin or flagellin-associatedpolypeptide and the amino acid sequences of the flagellin orflagellin-associated polypeptide comprises SEQ ID NO: 226, 540, 226,571, or 752, or any combination thereof and the RHPP polypeptide and theamino acid sequence of the RHPP polypeptide comprises SEQ ID NO: 600; or(i) the flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO: 226 and the RHPP polypeptide and the amino acid sequence ofthe RHPP polypeptide comprises SEQ ID NO:
 600. 25. The combination orrecombinant microorganism of any one of claims 12-24, further comprisingan assistance polypeptide.
 26. The composition or recombinantmicroorganism of claim 25 wherein the assistance polypeptide comprises:a signature polypeptide, and an amino acid sequence of the signaturepolypeptide comprises any one of SEQ ID NOs: 542-548, or any combinationthereof; or a signature polypeptide, and an amino acid sequence of thesignature polypeptide comprises SEQ ID NO: 542; or a signal anchorsorting polypeptide, and an amino acid sequence of the signal anchorsorting polypeptide comprises any one of SEQ ID NOs: 549-562, or anycombination thereof; or a signal anchor sorting polypeptide, and anamino acid sequence of the signal anchor sorting polypeptide comprisesany one of SEQ ID NOs: 549-562, or any combination thereof; or asecretion polypeptide, and an amino acid sequence of the secretionpolypeptide comprises any one of SEQ ID NOs: 563-570, or 769, or anycombination thereof; or a signature polypeptide, and an amino acidsequence of the signature polypeptide comprises SEQ ID NO: 542; or asignal anchor sorting polypeptide, and an amino acid sequence of thesignal anchor sorting polypeptide comprises SEQ ID NO: 549; or asecretion polypeptide, and an amino acid sequence of the secretionpolypeptide comprises SEQ ID NO: 563 or 769
 27. The composition orrecombinant microorganism of claim 26 wherein the assistance polypeptidefurther comprises: an enterokinase cleavage sequence and an amino acidsequence of the enterokinase cleavage sequence comprises SEQ ID NO: 772;or an enterokinase cleavage sequence and an amino acid sequence of theenterokinase cleavage sequence comprises SEQ ID NO: 772 and a linkingregion connecting the polypeptide and a protein tag; or an enterokinasecleavage sequence and an amino acid sequence of the enterokinasecleavage sequence comprises SEQ ID NO: 772 and a linking regionconnecting the polypeptide and a protein tag, wherein the protein tagoptionally comprises a poly-histidine (His) tag, a FLAG tag, an antibodyepitope, streptavidin/biotin, glutathione S-transferase (GST), or anycombination thereof.
 28. The composition of any one of claims 12-14 and18-27, further comprising an agrochemical or carrier which is associatedwith the polypeptide in nature.
 29. The composition of any one of claims12-14 and 18-28, wherein the agrochemical comprises an antibiotic, abiopesticide, a preservative, a buffering agent, a wetting agent, asurfactant, a coating agent, a monosaccharide, a polysaccharide, anabrading agent, a pesticide, an insecticide, a herbicide, a nematicide,a bacteriocide, a fungicide, a miticide, a fertilizer, a biostimulant,an osmoprotectant, a colorant, a humectant, an amino acid, a biologicalcontrol agent, hydrochloric acid, acetic acid, and/or trifluoroaceticacid or a combination thereof.
 30. The composition of any one of claims12-14 and 18-29, wherein the composition comprises: from about 0.00001wt. % to about 95 wt. % of the polypeptide(s), from about 0.01% to about80 wt. % of the agrochemical, and from about 5 wt. % to about 50 wt. %carrier based on the total weight of the composition; or from about 0.01wt. % to about 5 wt. % of the polypeptides, from about 0.2% to about 70wt. % of the agrochemical, and from about 10 wt. % to about 30 wt. %carrier based on the total weight of the composition; or from about 0.05wt. % to about 1 wt. % of the polypeptides, from about 30 wt,% to about60 wt. % of the agrochemical, and from about 40 wt. % to about 69 wt %carrier based on the total weight of the composition
 31. The compositionof claim 30 wherein the agrochemical comprises: hydrochloric acid,acetic acid, and/or trifluoroacetic acid; or about 0.001 wt. % to about30 wt. %, about 0.01 wt. % to about 20% wt. %, or 0.1 wt. % to about 5wt. % of hydrochloric acid, acetic acid, and/or trifluoroacetic acid,based on the total weight of the composition.
 32. A seed coated with thepolypeptide of any one of claims 1-12 or any combination thereof, thecomposition of any one of claims 12-14 and 18-31, or the recombinantmicroorganism of any one of claims 15 to
 27. 33. A method for increasinggrowth, yield, health, longevity, productivity, and/or vigor of a plantor a plant part and/or decreasing abiotic stress in the plant or theplant part and/or protecting the plant or the plant part from disease,insects and/or nematodes, and/or increasing the innate immune responseof the plant or the plant part and/or changing plant architecture, themethod comprising either: (a) applying the polypeptide of any one ofclaims 1-11 or the composition of any one of claims 12-14 and 18-31 to aplant, a plant part, or a plant growth medium or a rhizosphere in anarea surrounding the plant or the plant part to increase growth, yield,health, longevity, productivity, and/or vigor of the plant or the plantpart and/or decrease abiotic stress in the plant or the plant partand/or protect the plant or the plant part from disease, insects and/ornematodes, and/or increase the innate immune response of the plant orthe plant part and/or change the plant architecture; or (b) applying thepolypeptide of any one of claims 1-11 or the composition of any one ofclaims 12-14 and 18-31 to a plant growth medium to increase growth,yield, health, longevity, productivity, and/or vigor of a plant or aplant part to be grown in the plant growth medium and/or decreaseabiotic stress in the plant or the plant part to be grown in the plantgrowth medium and/or protect the plant or the plant part to be grown inthe plant growth medium from disease, insects and/or nematodes, and/orincrease the innate immune response and/or change plant architecture ofthe plant or the plant part to be grown in the plant growth medium; or(c) applying the recombinant microorganism of any one of claims 15-27 toa plant, a plant part, or a plant growth medium or a rhizosphere in anarea surrounding the plant or the plant part to increase growth, yield,health, longevity, productivity, and/or vigor of the plant or the plantpart and/or decrease abiotic stress in the plant or the plant partand/or protect the plant or the plant part from disease, insects and/ornematodes, and/or increase the innate immune response of the plant orthe plant part and/or change the plant architecture, wherein therecombinant microorganism expresses the polypeptide and expression ofthe polypeptide is increased as compared to the expression level thepolypeptide in a wild-type microorganism of the same kind under the sameconditions.
 34. The method of claim 33 wherein the method comprises: (A)decreasing abiotic stress in the plant or the plant part and/orprotecting the plant or the plant part from disease and/or increasingthe innate immune response of the plant or the plant part, and thepolypeptide or the composition comprises: the Flg22 polypeptide and anamino acid sequence of the Flg22 polypeptide comprises any one of SEQ IDNOs: 226-300 and 571-573 or any combination thereof; the retro inversoFlg22 polypeptide and an amino acid sequence of the retro inverso Flg22polypeptide comprises any one of SEQ ID NO: 376-450 or any combinationthereof; or any combination thereof; or the FlgII-28 polypeptide and anamino acid sequence of the FlgII-28 polypeptide comprises any one of SEQID NOs: 301-375 or any combination thereof; the retro inverso FlgII-28polypeptide and an amino acid sequence of the retro inverso FlgII-28polypeptide comprises any one of SEQ ID NO: 451-525 or any combinationthereof; or the retro inverso Flg22 polypeptide and an amino acidsequence of the retro inverso Flg22 polypeptide comprises any one of SEQID NO: 376-450 or any combination thereof; the retro inverso FlgII-28polypeptide and an amino acid sequence of the retro inverso FlgII-28polypeptide comprises any one of SEQ ID NO: 451-525 or any combinationthereof; or any combination thereof; the RHPP polypeptide and an aminoacid sequence of the RHPP polypeptide comprises SEQ ID NO: 600, 603, 604or any combination thereof; the Kunitz Trypsin Inhibitor (KTI)polypeptide and an amino acid sequence of the KTI polypeptide comprisesSEQ ID NO: 602; the retro-inverso RHPP polypeptide and an amino acidsequence of the RI RHPP comprises SEQ ID NO 601, 605, 606 or anycombination thereof or (B) protecting the plant or plant part fromdisease and/or increasing the innate immune response of the plant or theplant part, and the polypeptide or the composition comprises: (i) theFlg22 polypeptide and an amino acid sequence of the Flg22 polypeptidecomprises any one of SEQ ID NOs: 226-300 and 571-573; the retro inversoFlg22 polypeptide and an amino acid sequence of the retro inverso Flg22polypeptide comprises any one of SEQ ID NO: 376-450; or any combinationthereof; or (ii) the FlgII-28 polypeptide and an amino acid sequence ofthe FlgII-28 polypeptide comprises any one of SEQ ID NOs: 301-375; theretro inverso FlgII-28 polypeptide and an amino acid sequence of theretro inverso FlgII-28 polypeptide comprises any one of SEQ ID NO:451-525; or any combination thereof; or (iii) the Flg22 polypeptide andan amino acid sequence of the Flg22 polypeptide comprises any one of SEQID NOs: 226-300 and 571-573; the retro inverso Flg22 polypeptide and anamino acid sequence of the retro inverso Flg22 polypeptide comprises anyone of SEQ ID NO: 376-450 or any combination thereof; the FlgII-28polypeptide and an amino acid sequence of the FlgII-28 polypeptidecomprises any one of SEQ ID NOs: 301-375; the retro inverso FlgII-28polypeptide and an amino acid sequence of the retro inverso FlgII-28polypeptide comprises any one of SEQ ID NO: 451-525; or any combinationthereof; or (iv) the Flg22 polypeptide and an amino acid sequence of theFlg22 polypeptide comprises SEQ ID NO: 226, 752 or 571 and EF-Tupolypeptides and the amino acid sequences of the EF-Tu polypeptidescomprise SEQ ID NOs 616 and 617; or (C) protecting the plant or plantpart from disease, insects or nemotodes, and the polypeptide or thecomposition comprises: (i) the flagellin or flagellin-associatedpolypeptide and the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs: 226,289, 290, 291, 293, 294, 295, 300, 437, 532, 534, 536, 538, 540,571-586, and 751-766; or (ii) the flagellin or flagellin-associatedpolypeptide and the amino acid sequence of the flagellin orflagellin-associated polypeptide comprises any one of SEQ ID NOs: 226,293, 295, 300, 540, 571 574, 751, and 752 or any combination thereof; or(D) protecting the plant or plant part from disease, insects and/ornemotodes and/or increasing the innate immune response of the plant orthe plant part and the polypeptide or the composition comprises theharpin-like polypeptide or the RHPP polypeptide.
 35. The method of claim34 wherein either: (a) the method of (A) and when the polypeptide or thecomposition decreases abiotic stress, the abiotic stress comprises heatstress, temperature stress, radiation stress, drought stress, coldstress, salt stress, nutrient-deficient stress, high metal stress, waterstress, osmotic stress, or any combination thereof; or (b) the method of(B) or the method of (C) and the disease comprises Asian citrusgreening, Huanglonging (HLB) disease, Asian soybean rust, Sclerotiniastem rot (or white mold), Pseudomonas leaf spot, or Cercospora leafblight.
 36. The method of claim 33 wherein the method comprises: (A)increasing the yield, the growth, and/or the productivity of the plantor plant part and/or changing the plant architecture and the polypeptideor the composition comprises the RHPP polypeptide and/or the RI RHPPpolypeptide; or (B) increasing the yield of the plant or the plant partin environments prone to heat and drought and the polypeptide or thecomposition comprises the PSK polypeptide; or (C) increasing the growthof the plant or plant part and the polypeptide or the compositioncomprises the PSK polypeptide, the RHPP, the harpin or harpin-likepolypeptide, or a combination thereof; or (D) increasing the growth andproductivity of the plant or plant part in an environment prone to bothstress and non-stress conditions for plant growth and the polypeptide orthe composition comprises the PSK polypeptide and the harpin orharpin-like polypeptide
 37. The method of claim 36 wherein: (a) themethod of (A) and at least one of the following: (i) the RHPPpolypeptide of (A) comprises SEQ ID NO: 600; and/or (ii) the growth ofthe plant in (A) comprises root growth, root length, root biomass,nodulation, total biomass, above ground biomass, or any combinationthereof; and/or (iii) the plant in (A) comprises soybean, the growthcomprises overall root length, root biomass, nodulation, nodules perplant, total biomass, above ground biomass, or any combination thereof,and the productivity comprises number of total pods or pods per node;and/or (iv) the changes to the plant architecture in (A) comprisesbeneficial outcomes to the plant or plant part and/or (v) the changes tothe plant architecture in (A) comprises beneficial outcomes to the plantcomprising increased planting density capability for a field of theplants; or (b) the method of (B) and the polypeptide, composition orrecombinant microorganism of (B) is applied just prior to floralformation or at the pre-flowering stage; or (c) the method of (C) andthe growth of the plant of plant part in (C) comprises root and floralapical meristems, floral organ production, fruit development, fruitproduction, number of floral organs, size of floral organs, or acombination thereof; and/or
 38. The method of claim 35, wherein themethod of (B) or the method of (C) and the disease comprises Asiancitrus greening, Huanglonging (HLB) disease, Asian soybean rust,Sclerotinia stem rot (or white mold), Pseudomonas leaf spot, orCercospora, and the polypeptide or the composition comprises: the Flg22polypeptide and an amino acid sequence of the Flg22 polypeptidecomprises any one of SEQ ID NOs: 226-300 and 571-573 or any combinationthereof; or the FlgII-28 polypeptide and an amino acid sequence of theFlgII-28 polypeptide comprises any one of SEQ ID NOs: 301-375, or 751 orany combination thereof; or the Flg22 polypeptide and the FlgII-28polypeptide, an amino acid sequence of the Flg22 polypeptide comprisingany one of SEQ ID NOs: 226-300 and 571-573 or any combination thereofand an amino acid sequence of the FlgII-28 polypeptide comprising anyone of SEQ ID NOs: 301-375 or 751 or any combination thereof; the Flg22polypeptide and the FlgII-28 polypeptide, an amino acid sequence of theFlg22 polypeptide comprising any one of SEQ ID NOs: 226-300 and 571-573or any combination thereof and an amino acid sequence of the FlgII-28polypeptide comprising any one of SEQ ID NOs: 301-375 or 751 or anycombination thereof, and further comprising the retro inverso Flg22polypeptide, the retro inverso FlgII-28 polypeptide or a combinationthereof, an amino acid sequence of the retro inverso Flg22 polypeptidecomprising any one of SEQ ID NO: 376-450 or any combination thereof andan amino acid sequence of the retro inverso FlgII-28 polypeptidecomprising any one of SEQ ID NO: 451-525 or any combination thereof 39.The method of claim 33, wherein the disease comprises: (i) Asian soybeanrust, Cercospora leaf blight, or Scelerotinia stem rot (or white mold)and the polypeptide or the composition comprises: a flagellin orflagellin-associated polypeptide and the amino acid sequence of theflagellin or flagellin-associated polypeptide comprises any one of SEQID NOs: 226, 752, and 571; or an RHPP polypeptide and the amino acidsequence of the RHPP polypeptide comprises SEQ ID NO: 600; or aflagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 752, and 571 and an RHPP polypeptide and theamino acid sequence of the RHPP polypeptide comprises SEQ ID NO: 600; or(ii) Asian citrus greening and the composition comprises a bacteriocideand the flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 571, and 752 or any combination thereof; or(iii) Sclerotinia stem rot (or white mold) and the composition comprisesthe flagellin or flagellin-associated polypeptide and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs: 226, 540, 571, 751 and 752; or (iv) a bacterialdisease and wherein the method comprises restricting growth of thebacteria and/or preventing the disease and wherein the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesany one of SEQ ID NOs 226, 540, 752, and 571 or any combination thereof.40. The method of 39, wherein: the method of (i) and the amino acidsequence of the flagellin or flagellin-associated polypeptide comprisesSEQ ID NO 226; and the composition optionally comprises a fungicide orthe method of (iv) and the bacteria comprises Pseudomanas Syringae andthe plant, optionally, comprises a kiwi plant; OR the method of (ii) andthe bacteriocide comprises oxytetracycline.
 41. The method of claim 33,wherein the polypeptide or the composition comprises: the thionin orthionin-like polypeptide; or the thionin or thionin-like polypeptide orFlg22 polypeptides and the thionin or thionin-like polypeptide is fusedto a phloem targeting sequence to form a fused polypeptide, the aminoacid sequence of the phloem targeting sequence comprising any one of SEQID NOs: 641-649, or any combination thereof, for delivering the fusedpolypeptide to vascular tissue or cells and/or phloem orphloem-associated tissue or cells in the plant or plant part.
 42. Themethod of 33, wherein: the polypeptides comprise Flg22 polypeptides; orthe polypeptides comprise Flg22 polypeptides, and the Flg22 polypeptidescomprise SEQ ID NOs 526 and 527 or SEQ ID NOs 226 or 227 or thepolypeptide comprises the flagellin or flagellin-like polypeptide, andan amino acid sequence of the flagellin or flagellin-like polypeptidecomprising any one of SEQ ID NOs: 226-525 and 571-573 or any combinationthereof.
 43. The method of 41, wherein: (A) the disease comprises AsianCitrus disease (HLB), Citrus Canker disease, Cercospora leaf blight, ora bacteria causing disease; or (B) the disease comprises a bacteriacausing disease, and the bacteria causing disease comprises bacterialleaf blight, bacterial stalk rot, bacterial leaf spot, bacterial leafscorch, bacterial top rot, bacterial stripe, chocolate spot, Goss'sbacterial wilt and blight, Holcus spot, purple leaf sheath, seed rot,seedling blight, Stewart's disease (bacterial wilt), corn stunt, FireBlight, Pierce's disease, citrus variegated chlorosis, citrus canker,Pseudomonas syringae serovars, or a combination thereof.
 44. The methodof any one of claims 33 to 43, wherein protecting the plant or the plantpart from disease comprises prophylactic treatment, treatment,prevention and decreased disease progression on or in the plant or plantpart.
 45. The method of any one of claims 33 to 44, wherein thepolypeptide, the composition, or the recombinant microorganism isapplied exogenously to the plant, the plant part, or the plant growthmedium, or is applied endogenously to the plant or plant part.
 46. Themethod of any one of claims 33-45, wherein the plant part comprises acell, a leaf, a branch, a stem, a flower, a foliage, a floral organ, afruit, pollen, a vegetable, a tuber, a corm, a bulb, a pseudobulb, apod, a root, a root ball, a root stock, a scion, or a seed; and/or thepolypeptide, the composition, or the recombinant microorganism isapplied to a surface of the plant, a foliage of the plant or a surfaceof a seed of the plant; and/or the polypeptide, the composition, or therecombinant microorganism is applied to a surface of a seed of theplant, and the plant or the plant part is grown from the seed; and/orthe polypeptide, the composition, or the recombinant microorganism isapplied as a foliar application; and/or the plant is a fruit plant or avegetable plant and the method provides increased yield of fruits orvegetables.
 47. The method of producing a polypeptide, comprisingproducing a fusion protein comprising a polypeptide of any one of claims1 to 11, and an enterokinase (EK) cleavage site via fermentation, theenterokinase cleavage site enhancing activity and stability of thepolypeptide.
 48. The method of claim 47, wherein the method furthercomprises applying an enterokinase to the prepared fusion protein tocleave the enterokinase cleavage site and isolate the polypeptide. 49.The method of claim 47 or 48, wherein the enterokinase cleavage sitecomprises SEQ ID NO
 772. 50. The method of any one of claims 47-49,wherein: the fusion protein further comprises a protein tag; or thefusion protein further comprises a protein tag and the protein tagcomprises glutathione S transferase (GST); or the fusion protein furthercomprises a protein tag and a secretion signal; or the fusion proteinfurther comprises a protein tag and a secretion signal, and the proteintag comprises glutathione S transferase (GST); or the fusion proteinfurther comprises a protein tag and a secretion signal, and thesecretion signal comprises an amino acid sequence comprising any one ofSEQ ID NOs: 563-570, or 769, or any combination thereof; or the fusionprotein further comprises a protein tag and a secretion signal, and theprotein tag comprises glutathione S transferase (GST) and the secretionsignal comprises an amino acid sequence comprising any one of SEQ IDNOs: 563-570, or 769, or any combination thereof.